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Cell Signaling Technology Inc pparγ
(A) Activity of p-mTOR, SREBP-1, and <t>PPARγ</t> was decreased in supraspinatus muscles upon Rapamycin (Rap+) treatment as evident by Western blot analysis. No change was seen between vehicle (Rap-) and TT + DN groups. This data suggests there is a signaling relationship between mTOR, SREBP-1, and PPARγ in the setting of a RCT. (B) ImageJ quantitative analysis of Western blot bands ( * indicates p
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Images

1) Product Images from "mTOR Regulates Fatty Infiltration through SREBP-1 and PPARg after a Combined Massive Rotator Cuff Tear and Suprascapular Nerve Injury in Rats"

Article Title: mTOR Regulates Fatty Infiltration through SREBP-1 and PPARg after a Combined Massive Rotator Cuff Tear and Suprascapular Nerve Injury in Rats

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

doi: 10.1002/jor.22254

(A) Activity of p-mTOR, SREBP-1, and PPARγ was decreased in supraspinatus muscles upon Rapamycin (Rap+) treatment as evident by Western blot analysis. No change was seen between vehicle (Rap-) and TT + DN groups. This data suggests there is a signaling relationship between mTOR, SREBP-1, and PPARγ in the setting of a RCT. (B) ImageJ quantitative analysis of Western blot bands ( * indicates p
Figure Legend Snippet: (A) Activity of p-mTOR, SREBP-1, and PPARγ was decreased in supraspinatus muscles upon Rapamycin (Rap+) treatment as evident by Western blot analysis. No change was seen between vehicle (Rap-) and TT + DN groups. This data suggests there is a signaling relationship between mTOR, SREBP-1, and PPARγ in the setting of a RCT. (B) ImageJ quantitative analysis of Western blot bands ( * indicates p

Techniques Used: Activity Assay, Western Blot

ImageJ quantitative analysis of Western blot bands of Akt, p-Akt, mTOR, p-mTOR, SREBP-1, and PPARγ in supra-spinatus muscles 6 weeks after TT + DN surgery ( * indicates p
Figure Legend Snippet: ImageJ quantitative analysis of Western blot bands of Akt, p-Akt, mTOR, p-mTOR, SREBP-1, and PPARγ in supra-spinatus muscles 6 weeks after TT + DN surgery ( * indicates p

Techniques Used: Western Blot

Fold change of PPARγ, SREBP-1, C/EBPα, and FASN in supraspinatus muscles 6 weeks after TT + DN surgery ( * indicates p
Figure Legend Snippet: Fold change of PPARγ, SREBP-1, C/EBPα, and FASN in supraspinatus muscles 6 weeks after TT + DN surgery ( * indicates p

Techniques Used:

IHC of supraspinatus muscles 6 weeks after TT + DN surgery. There appears to be an up-regulation of p-mTOR (active), SREBP-1, and PPARγ in the surgical side (bottom row) compared to the sham control (top row).
Figure Legend Snippet: IHC of supraspinatus muscles 6 weeks after TT + DN surgery. There appears to be an up-regulation of p-mTOR (active), SREBP-1, and PPARγ in the surgical side (bottom row) compared to the sham control (top row).

Techniques Used: Immunohistochemistry

Fold change of PPARγ, SREBP-1, C/EBPα, and FASN in supraspinatus muscles 6 weeks after TT + DN surgery and rapamycin treatment ( * indicates p
Figure Legend Snippet: Fold change of PPARγ, SREBP-1, C/EBPα, and FASN in supraspinatus muscles 6 weeks after TT + DN surgery and rapamycin treatment ( * indicates p

Techniques Used:

Six weeks after TT + DN surgery, activity of Akt, p-Akt, mTOR, p-mTOR, SREBP-1, and PPARγ was up-regulated in supraspinatus muscles of the operated side compared to sham control as evident by Western blot analysis.
Figure Legend Snippet: Six weeks after TT + DN surgery, activity of Akt, p-Akt, mTOR, p-mTOR, SREBP-1, and PPARγ was up-regulated in supraspinatus muscles of the operated side compared to sham control as evident by Western blot analysis.

Techniques Used: Activity Assay, Western Blot

2) Product Images from "Mechanical Strain Downregulates C/EBP? in MSC and Decreases Endoplasmic Reticulum Stress"

Article Title: Mechanical Strain Downregulates C/EBP? in MSC and Decreases Endoplasmic Reticulum Stress

Journal: PLoS ONE

doi: 10.1371/journal.pone.0051613

C/EBPβ overexpression does not rescue adipogenesis from mechanical inhibition. A, C3H10T1/2 cells cultured in adipogenic media and analyzed on indicated days by real time PCR for C/EBPβ, PPARγ, and adiponectin (APN) n = 3, **p
Figure Legend Snippet: C/EBPβ overexpression does not rescue adipogenesis from mechanical inhibition. A, C3H10T1/2 cells cultured in adipogenic media and analyzed on indicated days by real time PCR for C/EBPβ, PPARγ, and adiponectin (APN) n = 3, **p

Techniques Used: Over Expression, Inhibition, Cell Culture, Real-time Polymerase Chain Reaction

3) Product Images from "TLE3 is a dual function transcriptional coregulator of adipogenesis"

Article Title: TLE3 is a dual function transcriptional coregulator of adipogenesis

Journal: Cell metabolism

doi: 10.1016/j.cmet.2011.02.014

TLE3 is a PPARγ target gene (A) Realtime PCR analysis of TLE mRNA expression in PPARγ2-expressing 10T1/2 preadipocytes treated with 100 nM GW7845 for 2 d (left) and 3T3-L1 preadipocytes treated with DMI + 20 nM GW for 2 d (right). (B) Induction of TLE3 and aP2 mRNA by PPARγ agonist in white and brown adipose tissue in vivo .
Figure Legend Snippet: TLE3 is a PPARγ target gene (A) Realtime PCR analysis of TLE mRNA expression in PPARγ2-expressing 10T1/2 preadipocytes treated with 100 nM GW7845 for 2 d (left) and 3T3-L1 preadipocytes treated with DMI + 20 nM GW for 2 d (right). (B) Induction of TLE3 and aP2 mRNA by PPARγ agonist in white and brown adipose tissue in vivo .

Techniques Used: Polymerase Chain Reaction, Expressing, In Vivo

TLE3 coactivates PPARγ-dependent gene expression .
Figure Legend Snippet: TLE3 coactivates PPARγ-dependent gene expression .

Techniques Used: Expressing

Overlapping transcriptional profiles of PPARγ and TLE3 regulated genes .
Figure Legend Snippet: Overlapping transcriptional profiles of PPARγ and TLE3 regulated genes .

Techniques Used:

4) Product Images from "Smooth Muscle Proliferation and Role of the Prostacyclin (IP) Receptor in Idiopathic Pulmonary Arterial Hypertension"

Article Title: Smooth Muscle Proliferation and Role of the Prostacyclin (IP) Receptor in Idiopathic Pulmonary Arterial Hypertension

Journal: American Journal of Respiratory and Critical Care Medicine

doi: 10.1164/rccm.201001-0011OC

Effect of peroxisome proliferator-activated receptor-gamma (PPARγ) modulators on cell growth. ( A ) Concentration-response curve to the PPARγ agonist rosiglitazone (ROSI) in human pulmonary arterial smooth muscle cells (PASMCs) and HEK-293-IP
Figure Legend Snippet: Effect of peroxisome proliferator-activated receptor-gamma (PPARγ) modulators on cell growth. ( A ) Concentration-response curve to the PPARγ agonist rosiglitazone (ROSI) in human pulmonary arterial smooth muscle cells (PASMCs) and HEK-293-IP

Techniques Used: Concentration Assay

( A ) Immunohistochemical staining for the IP receptor ( a , b , c ) and peroxisome proliferator-activated receptor-gamma (PPARγ) ( d , e , f ) in serial sections of pulmonary arteries from a normal and from an untreated or treated child with idiopathic
Figure Legend Snippet: ( A ) Immunohistochemical staining for the IP receptor ( a , b , c ) and peroxisome proliferator-activated receptor-gamma (PPARγ) ( d , e , f ) in serial sections of pulmonary arteries from a normal and from an untreated or treated child with idiopathic

Techniques Used: Immunohistochemistry, Staining

5) Product Images from "Neonatal diethylstilbestrol exposure alters the metabolic profile of uterine epithelial cells"

Article Title: Neonatal diethylstilbestrol exposure alters the metabolic profile of uterine epithelial cells

Journal: Disease Models & Mechanisms

doi: 10.1242/dmm.009076

DES affects lipid metabolism and transport. (A) Hierarchical clustering heatmap of genes involved in lipid trafficking and metabolism. Green to red, color range gradient of mean abundance (−1.0 to 1.0). Red box, genes whose expression was altered by DES primarily in the UE. (B) Real-time RT-PCR survey of genes involved in adipocyte differentiation in the uterus. Expression of each gene was normalized to that of Rpl7 ; normalized expression by oil UE was considered to be 1.0. (C) Western blot on whole uterine lysates showed markedly increased PPARγ protein in DES-treated uteri. GAPDH served as a loading control. (D,E) Immunohistochemistry of PPARγ showed increased staining in the DES-treated UE. Inset shows that PPARγ was predominantly detected in the UE nuclei (arrow). (F,G) Increased KLF4 protein was detected in the UE nuclei by immunofluorescence. Red, KLF4; blue, nuclei. Insets show magnified immunofluorescence signal of the boxed region. (H) Real-time RT-PCR validated genes involved in fatty acid transport and metabolism in oil- or DES-treated UE and UM. Data are presented as mean + s.d. of three samples analyzed in each treatment group from corresponding tissues. * P
Figure Legend Snippet: DES affects lipid metabolism and transport. (A) Hierarchical clustering heatmap of genes involved in lipid trafficking and metabolism. Green to red, color range gradient of mean abundance (−1.0 to 1.0). Red box, genes whose expression was altered by DES primarily in the UE. (B) Real-time RT-PCR survey of genes involved in adipocyte differentiation in the uterus. Expression of each gene was normalized to that of Rpl7 ; normalized expression by oil UE was considered to be 1.0. (C) Western blot on whole uterine lysates showed markedly increased PPARγ protein in DES-treated uteri. GAPDH served as a loading control. (D,E) Immunohistochemistry of PPARγ showed increased staining in the DES-treated UE. Inset shows that PPARγ was predominantly detected in the UE nuclei (arrow). (F,G) Increased KLF4 protein was detected in the UE nuclei by immunofluorescence. Red, KLF4; blue, nuclei. Insets show magnified immunofluorescence signal of the boxed region. (H) Real-time RT-PCR validated genes involved in fatty acid transport and metabolism in oil- or DES-treated UE and UM. Data are presented as mean + s.d. of three samples analyzed in each treatment group from corresponding tissues. * P

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Staining, Immunofluorescence

DES-induced lipogenesis in UE is mediated through PPAR γ. (A–D) TEM of UE at P5 revealed that DES treatment alone caused accumulation of electron-dense droplets in the UE close to the basement membrane (B, arrows, compare with A). Co-injection of PPARγ inhibitor BADGE (C) or GW9662 (D) dampened this effect, resulting in a lower number of droplets. (E–H) ORO staining identified droplets as neutral lipid droplets (F) and confirmed that co-treatment of DES with PPARγ inhibitors severely attenuated this effect (G,H). (I) Quantification of lipid droplets from TEM images confirmed DES-induced accumulation of lipid droplets in the UE. Co-treatment of DES with PPARγ inhibitors resulted in reduced number and size of the droplets. * P
Figure Legend Snippet: DES-induced lipogenesis in UE is mediated through PPAR γ. (A–D) TEM of UE at P5 revealed that DES treatment alone caused accumulation of electron-dense droplets in the UE close to the basement membrane (B, arrows, compare with A). Co-injection of PPARγ inhibitor BADGE (C) or GW9662 (D) dampened this effect, resulting in a lower number of droplets. (E–H) ORO staining identified droplets as neutral lipid droplets (F) and confirmed that co-treatment of DES with PPARγ inhibitors severely attenuated this effect (G,H). (I) Quantification of lipid droplets from TEM images confirmed DES-induced accumulation of lipid droplets in the UE. Co-treatment of DES with PPARγ inhibitors resulted in reduced number and size of the droplets. * P

Techniques Used: Transmission Electron Microscopy, Injection, Staining

6) Product Images from "Modulation of glycogen synthase kinase-3β following TRAIL combinatorial treatment in cancer cells"

Article Title: Modulation of glycogen synthase kinase-3β following TRAIL combinatorial treatment in cancer cells

Journal: Oncotarget

doi: 10.18632/oncotarget.11834

Effect of PPARγ knockdown on TRAIL-TZD-induced modulation of GSK3β pathway Subconfluent DU 145 cells were transfected with either control-siRNA or PPARγ-siRNA for 72 hrs followed by treatment with TRAIL or TZD alone or in combination for 6 hrs ( A ) or 24 hrs ( B ). The samples were analyzed by Western blots with the antibodies indicated.
Figure Legend Snippet: Effect of PPARγ knockdown on TRAIL-TZD-induced modulation of GSK3β pathway Subconfluent DU 145 cells were transfected with either control-siRNA or PPARγ-siRNA for 72 hrs followed by treatment with TRAIL or TZD alone or in combination for 6 hrs ( A ) or 24 hrs ( B ). The samples were analyzed by Western blots with the antibodies indicated.

Techniques Used: Transfection, Western Blot

7) Product Images from "Resolvin D1 attenuates inflammation in lipopolysaccharide-induced acute lung injury through a process involving the PPAR?/NF-?B pathway"

Article Title: Resolvin D1 attenuates inflammation in lipopolysaccharide-induced acute lung injury through a process involving the PPAR?/NF-?B pathway

Journal: Respiratory Research

doi: 10.1186/1465-9921-13-110

Effect of RvD1 on LPS-induced changes in expression of IκBα, NF-κB p-65 subunit and PPARγ in lung tissues. (a) Expression levels of IκBα, NF-κB p-65 and PPARγ. Histograms show mean ± S.D. (n = 3) of the relative intensity of (b) IκBα protein bands normalized to the β-actin band, (c) NF-κB p-65 protein bands normalized to the histone H3.1 band, and (d) PPARγ protein bands normalized to the histone H3.1 band. * p
Figure Legend Snippet: Effect of RvD1 on LPS-induced changes in expression of IκBα, NF-κB p-65 subunit and PPARγ in lung tissues. (a) Expression levels of IκBα, NF-κB p-65 and PPARγ. Histograms show mean ± S.D. (n = 3) of the relative intensity of (b) IκBα protein bands normalized to the β-actin band, (c) NF-κB p-65 protein bands normalized to the histone H3.1 band, and (d) PPARγ protein bands normalized to the histone H3.1 band. * p

Techniques Used: Expressing

8) Product Images from "Prokineticin receptor-1-dependent paracrine and autocrine pathways control cardiac tcf21+ fibroblast progenitor cell transformation into adipocytes and vascular cells"

Article Title: Prokineticin receptor-1-dependent paracrine and autocrine pathways control cardiac tcf21+ fibroblast progenitor cell transformation into adipocytes and vascular cells

Journal: Scientific Reports

doi: 10.1038/s41598-017-13198-2

Microarray analyses of TG-PKR1 hearts. ( A ) K-means analysis of the microarray data: gene expression is presented as a colored matrix, where each row represents a gene and each column represents a sample. Green, black and red correspond to lower values, median values and higher values, respectively. Left: transcriptome data clustered by K-means (with k = 10). The highlighted cluster 8 clearly distinguishes WT from TG-PKR1 mice (shown by hierarchical clustering on the right). ( B ) Hierarchical clustering of the samples based on the 46 expression profiles of WT and TG-PKR1 mice (12 weeks old). All n = 3 mice/group. ( C ) qPCR analyses of the PPARγ signaling pathway-related genes ( Fabp4 , UCP-1 , Scd1 , PPARγ , perilipin and adiponectin ) involved in adipogenesis (*p
Figure Legend Snippet: Microarray analyses of TG-PKR1 hearts. ( A ) K-means analysis of the microarray data: gene expression is presented as a colored matrix, where each row represents a gene and each column represents a sample. Green, black and red correspond to lower values, median values and higher values, respectively. Left: transcriptome data clustered by K-means (with k = 10). The highlighted cluster 8 clearly distinguishes WT from TG-PKR1 mice (shown by hierarchical clustering on the right). ( B ) Hierarchical clustering of the samples based on the 46 expression profiles of WT and TG-PKR1 mice (12 weeks old). All n = 3 mice/group. ( C ) qPCR analyses of the PPARγ signaling pathway-related genes ( Fabp4 , UCP-1 , Scd1 , PPARγ , perilipin and adiponectin ) involved in adipogenesis (*p

Techniques Used: Microarray, Expressing, Mouse Assay, Real-time Polymerase Chain Reaction

9) Product Images from "Licochalcone E protects against carbon tetrachloride-induced liver toxicity by activating peroxisome proliferator-activated receptor gamma"

Article Title: Licochalcone E protects against carbon tetrachloride-induced liver toxicity by activating peroxisome proliferator-activated receptor gamma

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2017.7268

Effect of LCE on PPARγ and NF-κB protein levels in mice liver tissue. (A) Measurement of PPARγ and NF-κB protein levels in mice liver were analyzed by western blot. Expression of NF-κB (B) and PPARγ (C) in tissue of mice liver by western blot analysis with β-actin was used as an internal control. ## P
Figure Legend Snippet: Effect of LCE on PPARγ and NF-κB protein levels in mice liver tissue. (A) Measurement of PPARγ and NF-κB protein levels in mice liver were analyzed by western blot. Expression of NF-κB (B) and PPARγ (C) in tissue of mice liver by western blot analysis with β-actin was used as an internal control. ## P

Techniques Used: Mouse Assay, Western Blot, Expressing

10) Product Images from "Brown Fat Determination and Development from Muscle Precursor Cells by Novel Action of Bone Morphogenetic Protein 6"

Article Title: Brown Fat Determination and Development from Muscle Precursor Cells by Novel Action of Bone Morphogenetic Protein 6

Journal: PLoS ONE

doi: 10.1371/journal.pone.0092608

BMP6 pretreatment induces temporal expression of Pparγ and commitment phase candidate genes. ( A ) Schematic representation of the BMP6 pretreatment segment of differentiation landscape illustrated in Figure 1A indicating the time points at which cells were harvested. ( B ) Heat map depiction and unsupervised clustering analyses of BMP6 target gene signature in cells stimulated with 250 ng/mL BMP6 and harvested at 0, 6, 12, 24 and 48 hours post BMP6 stimulation. Two way ANOVA was employed to extract 4046 significantly changing qualifiers ( p≤0.01 ) that were modulated as a function of treatment (BMP6 stimulation) and time (from 6 hours to 48 hours post BMP6 stimulation). Pparγ is highlighted in the heat map. Each gene is represented by a single row and each sample, in three independent biological replicates for each time point, by a column. Two distinct clusters indicate genes induced (red) and repressed (green). ( C ) Bar graph representation of Pparγ transcript levels at indicated time points measured using Affymetrix array. ( D ) Tabular representation of selected qualifiers, associated gene names and gene fold induction in cells stimulated with BMP6 relative to untreated cells at 12 hours. ( E ) Q-PCR validation of selected genes in cells stimulated with BMP6 for indicated time points. The expression in basal state (0 hour time point) was set to 1, and results represent triplicate analyses of three independent biological replicates (mean ± SD). Similar results were obtained in at least two independent analyses. Also see Figure S3 .
Figure Legend Snippet: BMP6 pretreatment induces temporal expression of Pparγ and commitment phase candidate genes. ( A ) Schematic representation of the BMP6 pretreatment segment of differentiation landscape illustrated in Figure 1A indicating the time points at which cells were harvested. ( B ) Heat map depiction and unsupervised clustering analyses of BMP6 target gene signature in cells stimulated with 250 ng/mL BMP6 and harvested at 0, 6, 12, 24 and 48 hours post BMP6 stimulation. Two way ANOVA was employed to extract 4046 significantly changing qualifiers ( p≤0.01 ) that were modulated as a function of treatment (BMP6 stimulation) and time (from 6 hours to 48 hours post BMP6 stimulation). Pparγ is highlighted in the heat map. Each gene is represented by a single row and each sample, in three independent biological replicates for each time point, by a column. Two distinct clusters indicate genes induced (red) and repressed (green). ( C ) Bar graph representation of Pparγ transcript levels at indicated time points measured using Affymetrix array. ( D ) Tabular representation of selected qualifiers, associated gene names and gene fold induction in cells stimulated with BMP6 relative to untreated cells at 12 hours. ( E ) Q-PCR validation of selected genes in cells stimulated with BMP6 for indicated time points. The expression in basal state (0 hour time point) was set to 1, and results represent triplicate analyses of three independent biological replicates (mean ± SD). Similar results were obtained in at least two independent analyses. Also see Figure S3 .

Techniques Used: Expressing, Polymerase Chain Reaction

11) Product Images from "Netrin-1-treated macrophages protect the kidney against ischemia-reperfusion injury and suppress inflammation by inducing M2 polarization"

Article Title: Netrin-1-treated macrophages protect the kidney against ischemia-reperfusion injury and suppress inflammation by inducing M2 polarization

Journal: American Journal of Physiology - Renal Physiology

doi: 10.1152/ajprenal.00580.2012

IFNγ-induced M1 marker expression and cytokine production are suppressed by netrin-1, and the effect of netrin-1 was abolished with PPAR antagonist. RAW264.7 cells were treated with netrin-1, IFNγ, netrin-1 + IFNγ, or PPARβ and -γ antagonist with netrin-1 and IFNγ for 48 h. Gene expression was analyzed by real-time PCR. IL-6 in the supernatant was quantified by ELISA. GW9662, PPARγ antagonist; GSK3797, PPARβ antagonist. Values are means ± SE ( n = 4). *Significant difference vs. vehicle-treated group ( P
Figure Legend Snippet: IFNγ-induced M1 marker expression and cytokine production are suppressed by netrin-1, and the effect of netrin-1 was abolished with PPAR antagonist. RAW264.7 cells were treated with netrin-1, IFNγ, netrin-1 + IFNγ, or PPARβ and -γ antagonist with netrin-1 and IFNγ for 48 h. Gene expression was analyzed by real-time PCR. IL-6 in the supernatant was quantified by ELISA. GW9662, PPARγ antagonist; GSK3797, PPARβ antagonist. Values are means ± SE ( n = 4). *Significant difference vs. vehicle-treated group ( P

Techniques Used: Marker, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

12) Product Images from "Peroxisome Proliferator-Activated Receptor γ Expression Is Inversely Associated with Macroscopic Vascular Invasion in Human Hepatocellular Carcinoma"

Article Title: Peroxisome Proliferator-Activated Receptor γ Expression Is Inversely Associated with Macroscopic Vascular Invasion in Human Hepatocellular Carcinoma

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms17081226

Peroxisome proliferator-activated receptor γ (PPARγ) and Krüppel-like factor 4 (KLF4) protein expression in human hepatocellular carcinoma (HCC) tissues: ( A ) representative views indicated PPARγ expression scores ranging from 0 to 3, as determined by immunohistochemistry (IHC); ( B ) the case number bar chart; and ( C ) correlation plot were generated using PPARγ and KLF4 staining scores from 83 human HCC tissue samples. Scale bar represents 100 μm.
Figure Legend Snippet: Peroxisome proliferator-activated receptor γ (PPARγ) and Krüppel-like factor 4 (KLF4) protein expression in human hepatocellular carcinoma (HCC) tissues: ( A ) representative views indicated PPARγ expression scores ranging from 0 to 3, as determined by immunohistochemistry (IHC); ( B ) the case number bar chart; and ( C ) correlation plot were generated using PPARγ and KLF4 staining scores from 83 human HCC tissue samples. Scale bar represents 100 μm.

Techniques Used: Expressing, Immunohistochemistry, Generated, Staining

Effects of PPARγ overexpression and knockdown on cell migration of Mahlavu cells and PLC/PRF/5 cells, respectively. ( A ) Cell migration abilities of Mahlavu-ctr and Mahlavu-PPARγ cells were analyzed over 14 h by wound healing assay; ( B ) cell migration abilities of PLC/PRF/5-shLuc and PLC/PRF/5-shPPARγ cells were assessed over 24 h by wound healing assay. Relative quantification data are expressed as the mean ± SEM (standard error of the mean) from three independent experiments. * p
Figure Legend Snippet: Effects of PPARγ overexpression and knockdown on cell migration of Mahlavu cells and PLC/PRF/5 cells, respectively. ( A ) Cell migration abilities of Mahlavu-ctr and Mahlavu-PPARγ cells were analyzed over 14 h by wound healing assay; ( B ) cell migration abilities of PLC/PRF/5-shLuc and PLC/PRF/5-shPPARγ cells were assessed over 24 h by wound healing assay. Relative quantification data are expressed as the mean ± SEM (standard error of the mean) from three independent experiments. * p

Techniques Used: Over Expression, Migration, Planar Chromatography, Wound Healing Assay

Effects of PPARγ overexpression and knockdown on in vitro human umbilical vein endothelial cells (HUVEC) tube formation in Mahlavu and PLC/PRF/5 cells, respectively. Conditioned medium was harvested from: ( A ) Mahlavu-ctr and Mahlavu-PPARγ; and ( B ) PLC/PRF/5-shLuc and PLC/PRF/5-shPPARγ cell cultures. The conditioned medium was used for HUVEC cell tube formation, and photos were taken after 6 h of incubation. Quantification data are expressed as the mean ± SEM from three independent experiments. * p
Figure Legend Snippet: Effects of PPARγ overexpression and knockdown on in vitro human umbilical vein endothelial cells (HUVEC) tube formation in Mahlavu and PLC/PRF/5 cells, respectively. Conditioned medium was harvested from: ( A ) Mahlavu-ctr and Mahlavu-PPARγ; and ( B ) PLC/PRF/5-shLuc and PLC/PRF/5-shPPARγ cell cultures. The conditioned medium was used for HUVEC cell tube formation, and photos were taken after 6 h of incubation. Quantification data are expressed as the mean ± SEM from three independent experiments. * p

Techniques Used: Over Expression, In Vitro, Planar Chromatography, Incubation

Effects of PPARγ overexpression and knockdown on cell proliferation and PPARγ downstream target protein expression in Mahlavu and PLC/PRF/5 HCC cells, respectively. ( A , C ) The cell proliferation rates of Mahlavu-ctr, Mahlavu-PPARγ, PLC/PRF/5-shLuc, and PLC/PRF/5-shPPARγ cells were analyzed by SRB assay; ( B , D ) the expression of PPARγ downstream target proteins STAT3 and cyclin D1 was analyzed by Western blot and the quantification results are shown. * p
Figure Legend Snippet: Effects of PPARγ overexpression and knockdown on cell proliferation and PPARγ downstream target protein expression in Mahlavu and PLC/PRF/5 HCC cells, respectively. ( A , C ) The cell proliferation rates of Mahlavu-ctr, Mahlavu-PPARγ, PLC/PRF/5-shLuc, and PLC/PRF/5-shPPARγ cells were analyzed by SRB assay; ( B , D ) the expression of PPARγ downstream target proteins STAT3 and cyclin D1 was analyzed by Western blot and the quantification results are shown. * p

Techniques Used: Over Expression, Expressing, Planar Chromatography, Sulforhodamine B Assay, Western Blot

13) Product Images from "Regulation of steatohepatitis and PPARγ signaling by distinct AP-1 dimers"

Article Title: Regulation of steatohepatitis and PPARγ signaling by distinct AP-1 dimers

Journal: Cell metabolism

doi: 10.1016/j.cmet.2013.11.018

Antagonistic regulation of PPARγ signaling by AP-1
Figure Legend Snippet: Antagonistic regulation of PPARγ signaling by AP-1

Techniques Used:

Fra-1 regulates the PPARγ pathway
Figure Legend Snippet: Fra-1 regulates the PPARγ pathway

Techniques Used:

Several AP-1 proteins regulate the PPARγ pathway
Figure Legend Snippet: Several AP-1 proteins regulate the PPARγ pathway

Techniques Used:

Fra-1 is regulated by HFD and inhibits NAFLD and PPARγ expression
Figure Legend Snippet: Fra-1 is regulated by HFD and inhibits NAFLD and PPARγ expression

Techniques Used: Expressing

PPARγ delivery restores NAFLD development in Fra-1 hep mice
Figure Legend Snippet: PPARγ delivery restores NAFLD development in Fra-1 hep mice

Techniques Used: Mouse Assay

14) Product Images from "Cyclic AMP Mimics the Anti-ageing Effects of Calorie Restriction by Up-Regulating Sirtuin"

Article Title: Cyclic AMP Mimics the Anti-ageing Effects of Calorie Restriction by Up-Regulating Sirtuin

Journal: Scientific Reports

doi: 10.1038/srep12012

cAMP Treatment Improves Adipose Metabolism to Mimic Calorie Restriction. ( A ) The curve for the body weight gain. ( B ) Daily food intake. ( C ) Latency to fall from a rota-rod indicated the motor coordination of mice. ( D ) The photographs of epididymal adipose tissue. ( E ) Histological analysis of liver tissue, which was stained with oil red O to visualize lipid content and counterstained with haematoxylin. ( F ) Abdominal adipose tissue stained with haematoxylin and eosin. ( G ) The total cholesterol in mouse liver. ( H ) The total glycerides in mouse liver. ( I ) Western blot results for the PKAc, PEPCK, PPARγ, CaMKKII, SIRT1 and SIRT3 protein levels in treated or untreated mouse liver. The gels were run under the same experimental conditions. ( J ) Fold change in Adiponectin levels in liver. ( K ) MDA level in mice liver. All the statistical analyses were performed using one-way ANOVA. # p
Figure Legend Snippet: cAMP Treatment Improves Adipose Metabolism to Mimic Calorie Restriction. ( A ) The curve for the body weight gain. ( B ) Daily food intake. ( C ) Latency to fall from a rota-rod indicated the motor coordination of mice. ( D ) The photographs of epididymal adipose tissue. ( E ) Histological analysis of liver tissue, which was stained with oil red O to visualize lipid content and counterstained with haematoxylin. ( F ) Abdominal adipose tissue stained with haematoxylin and eosin. ( G ) The total cholesterol in mouse liver. ( H ) The total glycerides in mouse liver. ( I ) Western blot results for the PKAc, PEPCK, PPARγ, CaMKKII, SIRT1 and SIRT3 protein levels in treated or untreated mouse liver. The gels were run under the same experimental conditions. ( J ) Fold change in Adiponectin levels in liver. ( K ) MDA level in mice liver. All the statistical analyses were performed using one-way ANOVA. # p

Techniques Used: Mouse Assay, Staining, Western Blot, Multiple Displacement Amplification

15) Product Images from "Epoxyeicosatrienoic acids protect rat hearts against tumor necrosis factor-?-induced injury [S]"

Article Title: Epoxyeicosatrienoic acids protect rat hearts against tumor necrosis factor-?-induced injury [S]

Journal: Journal of Lipid Research

doi: 10.1194/jlr.M017319

Effect of CYP2J2 overexpression and 11,12-EET treatment on PPARγ and IκBα expression in neonatal rat cardiac myocytes. (A) Representative Western blots and densitometry results showing effects of CYP2J2 overexpression on PPARγ
Figure Legend Snippet: Effect of CYP2J2 overexpression and 11,12-EET treatment on PPARγ and IκBα expression in neonatal rat cardiac myocytes. (A) Representative Western blots and densitometry results showing effects of CYP2J2 overexpression on PPARγ

Techniques Used: Over Expression, Expressing, Western Blot

16) Product Images from "Annual Wormwood Leaf Inhibits the Adipogenesis of 3T3-L1 and Obesity in High-Fat Diet-Induced Obese Rats"

Article Title: Annual Wormwood Leaf Inhibits the Adipogenesis of 3T3-L1 and Obesity in High-Fat Diet-Induced Obese Rats

Journal: Nutrients

doi: 10.3390/nu9060554

Effects of AWL on mRNA expression in epididymal adipose tissues. AWL decreased the expression of adipogenic factors in adipocytes. PPARγ, peroximal proliferator-activated receptor-γ; C/EPBα, CCAAT/enhancer binding protein-α; ACC, acetyl-CoA carboxylase; aP2, adipocyte fatty acid-binding protein 4; FAS, fatty acid synthase. The values are presented as the means ± SD. The data shown are representative of at least three independent experiments. β-actin expression in each sample was used as an internal control to normalize expression. * p
Figure Legend Snippet: Effects of AWL on mRNA expression in epididymal adipose tissues. AWL decreased the expression of adipogenic factors in adipocytes. PPARγ, peroximal proliferator-activated receptor-γ; C/EPBα, CCAAT/enhancer binding protein-α; ACC, acetyl-CoA carboxylase; aP2, adipocyte fatty acid-binding protein 4; FAS, fatty acid synthase. The values are presented as the means ± SD. The data shown are representative of at least three independent experiments. β-actin expression in each sample was used as an internal control to normalize expression. * p

Techniques Used: Expressing, Binding Assay

17) Product Images from "Rosehip Extract Inhibits Lipid Accumulation in White Adipose Tissue by Suppressing the Expression of Peroxisome Proliferator-activated Receptor Gamma"

Article Title: Rosehip Extract Inhibits Lipid Accumulation in White Adipose Tissue by Suppressing the Expression of Peroxisome Proliferator-activated Receptor Gamma

Journal: Preventive Nutrition and Food Science

doi: 10.3746/pnf.2013.18.2.085

Effect of RHE on PPARγ expression in epididymal fat. Male C57BL/6 J mice were fed HFD or HFDRH for 8 weeks. On the final day, a part of epididymal fat was collected and total protein extracted. The amount of PPARγ was quantified by Western blotting. Protein expression of PPARγ was quantified and normalized against GAPDH. Each column represents mean±SEM and n=5. Significantly different from the HFD-fed group (white column), * P
Figure Legend Snippet: Effect of RHE on PPARγ expression in epididymal fat. Male C57BL/6 J mice were fed HFD or HFDRH for 8 weeks. On the final day, a part of epididymal fat was collected and total protein extracted. The amount of PPARγ was quantified by Western blotting. Protein expression of PPARγ was quantified and normalized against GAPDH. Each column represents mean±SEM and n=5. Significantly different from the HFD-fed group (white column), * P

Techniques Used: Expressing, Mouse Assay, Western Blot

18) Product Images from "Differential transcription factor expression by human epithelial cells of buccal and urothelial derivation"

Article Title: Differential transcription factor expression by human epithelial cells of buccal and urothelial derivation

Journal: Experimental Cell Research

doi: 10.1016/j.yexcr.2018.05.031

Evaluation of PPARγ, FOXA1 and GATA3 expression in PPARG1 overexpressing and control (empty) NHB cells. PPARG1 overexpressing and control (empty vector) NHB cell cultures were exposed to the TZ/PD protocol for 72 h. Experiments performed on between 2 and 4 independent NHB cell lines (as stated below), with representative results shown. (A) PPARγ1, FOXA1 and GATA3 protein expression assessed by western blotting. ACTB expression was included as an internal loading control. Protein lysates from cell lines known to express the proteins of interest were included as positive controls for each antibody (CTRL). Experiments performed on n = 3 independent NHB cell lines. (B) Densitometry analysis of FOXA1 protein expression shown relative to control (Empty - DMSO) NHB cells. Data is shown as the mean of n = 4 independent transduced NHB cell lines. All values were normalised to the ACTB expression. Statistical analysis was performed using a one-way ANOVA test, but no statistical significance was found (P > 0.05). Error bars represent standard deviation. (C) Immunofluorescence microscopy evaluating PPARγ, FOXA1 and GATA3 protein localisation in PPARG1 overexpressing and control (empty vector) NHB cells following the TZ/PD protocol at 72 h. Experiments were performed on n = 3 independent transduced NHB cell lines. IF images for a single NHB cell line are shown. Nuclear localisation was observed with PPARγ (n = 3/3), FOXA1 (n = 3/3), and GATA3 (n = 2/3). NHU cells (non-transduced) treated with the TZ/PD protocol are shown for comparison at the same time point. Scale bar ≡ 50 µm.
Figure Legend Snippet: Evaluation of PPARγ, FOXA1 and GATA3 expression in PPARG1 overexpressing and control (empty) NHB cells. PPARG1 overexpressing and control (empty vector) NHB cell cultures were exposed to the TZ/PD protocol for 72 h. Experiments performed on between 2 and 4 independent NHB cell lines (as stated below), with representative results shown. (A) PPARγ1, FOXA1 and GATA3 protein expression assessed by western blotting. ACTB expression was included as an internal loading control. Protein lysates from cell lines known to express the proteins of interest were included as positive controls for each antibody (CTRL). Experiments performed on n = 3 independent NHB cell lines. (B) Densitometry analysis of FOXA1 protein expression shown relative to control (Empty - DMSO) NHB cells. Data is shown as the mean of n = 4 independent transduced NHB cell lines. All values were normalised to the ACTB expression. Statistical analysis was performed using a one-way ANOVA test, but no statistical significance was found (P > 0.05). Error bars represent standard deviation. (C) Immunofluorescence microscopy evaluating PPARγ, FOXA1 and GATA3 protein localisation in PPARG1 overexpressing and control (empty vector) NHB cells following the TZ/PD protocol at 72 h. Experiments were performed on n = 3 independent transduced NHB cell lines. IF images for a single NHB cell line are shown. Nuclear localisation was observed with PPARγ (n = 3/3), FOXA1 (n = 3/3), and GATA3 (n = 2/3). NHU cells (non-transduced) treated with the TZ/PD protocol are shown for comparison at the same time point. Scale bar ≡ 50 µm.

Techniques Used: Expressing, Plasmid Preparation, Western Blot, Standard Deviation, Immunofluorescence, Microscopy

GATA3 overexpression in NHB cells. (A) GATA3 overexpressing and control (empty vector) NHB cell cultures following exposure to the PPARγ-activating TZ/PD protocol for 72 h. Western blotting of whole protein lysates was performed to assess protein expression of GATA3, FOXA1 and PPARy1. NHU cells (non-transduced) and treated with the TZ/PD protocol for 72 h are shown for comparison. (B) GATA3 overexpressing and control (empty vector) NHB cells at 72 h post TZ/PD protocol. GATA3, FOXA1 and PPARy protein expression assessed by indirect immunofluorescence microscopy. NHU cells (non-transduced; 72 h TZ/PD protocol) were included as positive controls for comparison. Scale bar ≡ 50 µm. (C) GATA3 overexpressing and control (empty vector) NHB cells were induced to form cell sheets using 5% ABS and 2 mM calcium for up to 7 days. Expression of the tight junction-associated proteins, claudin 3, 4, 5 and 7, assessed by western blotting. ACTB was included as a loading control. NHU cells (non-transduced) exposed to the same protocol were used as a positive control for comparison. Experiments were performed on n = 2 independent NHB donor cell lines and representative results shown.
Figure Legend Snippet: GATA3 overexpression in NHB cells. (A) GATA3 overexpressing and control (empty vector) NHB cell cultures following exposure to the PPARγ-activating TZ/PD protocol for 72 h. Western blotting of whole protein lysates was performed to assess protein expression of GATA3, FOXA1 and PPARy1. NHU cells (non-transduced) and treated with the TZ/PD protocol for 72 h are shown for comparison. (B) GATA3 overexpressing and control (empty vector) NHB cells at 72 h post TZ/PD protocol. GATA3, FOXA1 and PPARy protein expression assessed by indirect immunofluorescence microscopy. NHU cells (non-transduced; 72 h TZ/PD protocol) were included as positive controls for comparison. Scale bar ≡ 50 µm. (C) GATA3 overexpressing and control (empty vector) NHB cells were induced to form cell sheets using 5% ABS and 2 mM calcium for up to 7 days. Expression of the tight junction-associated proteins, claudin 3, 4, 5 and 7, assessed by western blotting. ACTB was included as a loading control. NHU cells (non-transduced) exposed to the same protocol were used as a positive control for comparison. Experiments were performed on n = 2 independent NHB donor cell lines and representative results shown.

Techniques Used: Over Expression, Plasmid Preparation, Western Blot, Expressing, Immunofluorescence, Microscopy, Positive Control

Comparison of uroplakin and urothelium differentiation-associated transcription factor gene expression by NHB and NHU cell cultures. Employing protocols developed to differentiate NHU cells by PPARγ activation, cell cultures of NHB or NHU cells were exposed to 1 µM troglitazone and 1 µM PD153035 (TZ/PD) for 24 h, maintained in 1 µM PD153035 and harvested at 12, 24, 48 and/or 72 h. Control cultures were exposed to vehicle (0.1% DMSO) alone. (A) RTqPCR for three independent NHB cell lines (represented by different symbols), versus a single NHU cell line for comparison of uroplakin (UPK1A, UPK1B, UPK2, UPK3A and UPK3B) mRNA expression at the 72 h time-point. All data has been normalised to GAPDH expression and is presented relative to the DMSO-treated NHB cells for each gene except UPK3A, where the data is shown relative to the DMSO treated NHU cells due to absent UPK3A gene expression by NHB cells. BLOD = Below Limit of Detection. Statistical analysis was performed using a two-tailed, paired t -test to determine whether TZ/PD resulted in any significant change in gene expression in NHB cells. *represents P ≤ 0.05, ** represents P ≤ 0.01. Error bars represent standard deviation. (B-C) RT-PCR of ELF3, FOXA1, GATA3, GRHL3, IRF1, KLF5 and PPARG mRNA expression by (B) NHB cells and (C) NHU cells. RNA was extracted at the 12, 24 and 48 h time-points and then DNAase-treated and used to generate cDNA for RT-PCR. GAPDH was used as an internal loading control. A no-template (H 2 O) control was included as a negative control for the PCR reaction, and genomic DNA was used as the positive control (+ctrl). No product was amplified from RT-negative controls (not shown). Experiments were performed on n = 2 independent NHB donor cell lines and representative results shown.
Figure Legend Snippet: Comparison of uroplakin and urothelium differentiation-associated transcription factor gene expression by NHB and NHU cell cultures. Employing protocols developed to differentiate NHU cells by PPARγ activation, cell cultures of NHB or NHU cells were exposed to 1 µM troglitazone and 1 µM PD153035 (TZ/PD) for 24 h, maintained in 1 µM PD153035 and harvested at 12, 24, 48 and/or 72 h. Control cultures were exposed to vehicle (0.1% DMSO) alone. (A) RTqPCR for three independent NHB cell lines (represented by different symbols), versus a single NHU cell line for comparison of uroplakin (UPK1A, UPK1B, UPK2, UPK3A and UPK3B) mRNA expression at the 72 h time-point. All data has been normalised to GAPDH expression and is presented relative to the DMSO-treated NHB cells for each gene except UPK3A, where the data is shown relative to the DMSO treated NHU cells due to absent UPK3A gene expression by NHB cells. BLOD = Below Limit of Detection. Statistical analysis was performed using a two-tailed, paired t -test to determine whether TZ/PD resulted in any significant change in gene expression in NHB cells. *represents P ≤ 0.05, ** represents P ≤ 0.01. Error bars represent standard deviation. (B-C) RT-PCR of ELF3, FOXA1, GATA3, GRHL3, IRF1, KLF5 and PPARG mRNA expression by (B) NHB cells and (C) NHU cells. RNA was extracted at the 12, 24 and 48 h time-points and then DNAase-treated and used to generate cDNA for RT-PCR. GAPDH was used as an internal loading control. A no-template (H 2 O) control was included as a negative control for the PCR reaction, and genomic DNA was used as the positive control (+ctrl). No product was amplified from RT-negative controls (not shown). Experiments were performed on n = 2 independent NHB donor cell lines and representative results shown.

Techniques Used: Expressing, Activation Assay, Two Tailed Test, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Negative Control, Polymerase Chain Reaction, Positive Control, Amplification

Evaluation of ELF3 , FOXA1 , GATA3 and PPARG expression in NHB cells. RNA and protein were extracted from parallel cultures of NHB and NHU cells at 72 h following exposure to the PPARγ-activating TZ/PD protocol, or a vehicle control (0.1% DMSO). (A) RTqPCR results combined from three independent NHB cell lines (represented by different symbols), with a single NHU cell line for comparison. All data is normalised to GAPDH expression and is presented relative to the DMSO-treated NHB cell control for each gene. Statistical analysis was performed using a two-tailed, paired t -test to test if TZ/PD treatment resulted in any significant change in gene expression in NHB cells. ** represents P ≤ 0.01. Error bars represent standard deviation. (B) Immunoblot of whole cell protein lysates from representative NHB and NHU cell cultures following exposure to the TZ/PD protocol, 1 µM PD153035 alone, or vehicle (0.1% DMSO) for 72 h. ACTB was included as an internal loading control. Experiments performed on n = 3 independent NHB cell lines with similar results. (C) Immunofluorescence microscopy of ELF3, FOXA1/2, GATA3 and PPARγ in representative NHB and NHU cell cultures. Images taken at identical exposures to demonstrate differences in labelling intensity between the two cell types. Experiments performed on n = 3 independent NHB donor cell lines with similar results. Scale bar ≡ 50 µm.
Figure Legend Snippet: Evaluation of ELF3 , FOXA1 , GATA3 and PPARG expression in NHB cells. RNA and protein were extracted from parallel cultures of NHB and NHU cells at 72 h following exposure to the PPARγ-activating TZ/PD protocol, or a vehicle control (0.1% DMSO). (A) RTqPCR results combined from three independent NHB cell lines (represented by different symbols), with a single NHU cell line for comparison. All data is normalised to GAPDH expression and is presented relative to the DMSO-treated NHB cell control for each gene. Statistical analysis was performed using a two-tailed, paired t -test to test if TZ/PD treatment resulted in any significant change in gene expression in NHB cells. ** represents P ≤ 0.01. Error bars represent standard deviation. (B) Immunoblot of whole cell protein lysates from representative NHB and NHU cell cultures following exposure to the TZ/PD protocol, 1 µM PD153035 alone, or vehicle (0.1% DMSO) for 72 h. ACTB was included as an internal loading control. Experiments performed on n = 3 independent NHB cell lines with similar results. (C) Immunofluorescence microscopy of ELF3, FOXA1/2, GATA3 and PPARγ in representative NHB and NHU cell cultures. Images taken at identical exposures to demonstrate differences in labelling intensity between the two cell types. Experiments performed on n = 3 independent NHB donor cell lines with similar results. Scale bar ≡ 50 µm.

Techniques Used: Expressing, Two Tailed Test, Standard Deviation, Immunofluorescence, Microscopy

19) Product Images from "Identification of Cells with Colony-Forming Activity, Self-Renewal Capacity, and Multipotency in Ovarian Endometriosis"

Article Title: Identification of Cells with Colony-Forming Activity, Self-Renewal Capacity, and Multipotency in Ovarian Endometriosis

Journal: The American Journal of Pathology

doi: 10.1016/j.ajpath.2011.02.025

Multipotency of endometriotic stromal CFUs. In vitro differentiation of endometriotic stromal CFUs toward the adipogenic ( A–C ), myogenic ( D–E ), osteogenic ( F–G ), and chondrogenic (H–J) lineages was confirmed with histochemical staining, IHC, and RT-PCR by mRNA expression of specific lineage markers. A : Adipogenic differentiation with fluorescent lipid stain (green) on cells clonally derived from endometriotic stromal large CFUs. B : Positive staining on mouse adipose tissue. C : IHC staining of peroxisome proliferation activated receptor γ (PPARγ) and expression of PPARG, CEBPA (CCAAT/enhanced binding protein alpha) and LPL (lipoprotein lipase). Myogenic differentiation with α smooth muscle cell marker (α-SMA, brown) staining on cells clonally derived from endometriotic stromal large CFUs ( D ) and untreated endometriotic stromal cells ( E ). F and G : Expression of ACTA2 (encoding α-SMA), caldesmon (CALD1) and CNN1 (calponin 1). Osteogenic differentiation with alkaline phosphates (red) staining ( F ) and IHC staining of osteopontin ( G ) and expression of alkaline phosphates (ALPL), RUNX2 (runt-related transcription factor 2), SSP1 (secreted phosphoprotein 1, previously OPN, osteopontin), and parathyroid hormone 1 receptor (PTH1R). H–J : Chondrogenic differentiation shown in paraffin section of micropellet stained with Alcian Blue ( H ) and Safranin O (red, arrows ) ( I ); IHC staining of collagen type II ( J ). Expression of COL2A1 (collagen, type II, alpha 1) and COL10A1 (collagen, type X, alpha 1). Cells cultured in control culture medium for 4 weeks and stained for lineage markers are shown in insets ( F , G , and I ) and as negative control (−) for RT-PCR analysis; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA was the internal control. Results shown are from a single sample representative of three patients. Scale bars = 200 μm.
Figure Legend Snippet: Multipotency of endometriotic stromal CFUs. In vitro differentiation of endometriotic stromal CFUs toward the adipogenic ( A–C ), myogenic ( D–E ), osteogenic ( F–G ), and chondrogenic (H–J) lineages was confirmed with histochemical staining, IHC, and RT-PCR by mRNA expression of specific lineage markers. A : Adipogenic differentiation with fluorescent lipid stain (green) on cells clonally derived from endometriotic stromal large CFUs. B : Positive staining on mouse adipose tissue. C : IHC staining of peroxisome proliferation activated receptor γ (PPARγ) and expression of PPARG, CEBPA (CCAAT/enhanced binding protein alpha) and LPL (lipoprotein lipase). Myogenic differentiation with α smooth muscle cell marker (α-SMA, brown) staining on cells clonally derived from endometriotic stromal large CFUs ( D ) and untreated endometriotic stromal cells ( E ). F and G : Expression of ACTA2 (encoding α-SMA), caldesmon (CALD1) and CNN1 (calponin 1). Osteogenic differentiation with alkaline phosphates (red) staining ( F ) and IHC staining of osteopontin ( G ) and expression of alkaline phosphates (ALPL), RUNX2 (runt-related transcription factor 2), SSP1 (secreted phosphoprotein 1, previously OPN, osteopontin), and parathyroid hormone 1 receptor (PTH1R). H–J : Chondrogenic differentiation shown in paraffin section of micropellet stained with Alcian Blue ( H ) and Safranin O (red, arrows ) ( I ); IHC staining of collagen type II ( J ). Expression of COL2A1 (collagen, type II, alpha 1) and COL10A1 (collagen, type X, alpha 1). Cells cultured in control culture medium for 4 weeks and stained for lineage markers are shown in insets ( F , G , and I ) and as negative control (−) for RT-PCR analysis; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA was the internal control. Results shown are from a single sample representative of three patients. Scale bars = 200 μm.

Techniques Used: In Vitro, Staining, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Expressing, Derivative Assay, Binding Assay, Marker, Paraffin Section, Cell Culture, Negative Control

20) Product Images from "Women Have Higher Protein Content of ?-Oxidation Enzymes in Skeletal Muscle than Men"

Article Title: Women Have Higher Protein Content of ?-Oxidation Enzymes in Skeletal Muscle than Men

Journal: PLoS ONE

doi: 10.1371/journal.pone.0012025

No sex differences in PPARα or PPARγ protein content. Protein content of PPARα (A) and PPARγ (B) in skeletal muscle of men and women, adjusted to actin. Representative western blots; lanes 1–4 are men, lanes 5–8 women. N = 12 men and 11 women.
Figure Legend Snippet: No sex differences in PPARα or PPARγ protein content. Protein content of PPARα (A) and PPARγ (B) in skeletal muscle of men and women, adjusted to actin. Representative western blots; lanes 1–4 are men, lanes 5–8 women. N = 12 men and 11 women.

Techniques Used: Western Blot

21) Product Images from "TET2 facilitates PPARγ agonist–mediated gene regulation and insulin sensitization in adipocytes"

Article Title: TET2 facilitates PPARγ agonist–mediated gene regulation and insulin sensitization in adipocytes

Journal: Metabolism: clinical and experimental

doi: 10.1016/j.metabol.2018.08.006

TET2 facilitates the transcriptional activity of PPARγ via physical interaction. ( A, B ) 293T cells were co-transfected with vectors expressing Tet2-CD, Tet2-HxD, PPARγ, and reporter plasmids containing 3 copies of PPREs ( A ) and 2.4 kb of the Glut4 promoter ( B ). We measured Luciferase and Renilla activity 24 hr after transfection. Shown is the relative fold stimulation of luciferase activity after assessing transfection efficiency with Renilla ( n = 3, p
Figure Legend Snippet: TET2 facilitates the transcriptional activity of PPARγ via physical interaction. ( A, B ) 293T cells were co-transfected with vectors expressing Tet2-CD, Tet2-HxD, PPARγ, and reporter plasmids containing 3 copies of PPREs ( A ) and 2.4 kb of the Glut4 promoter ( B ). We measured Luciferase and Renilla activity 24 hr after transfection. Shown is the relative fold stimulation of luciferase activity after assessing transfection efficiency with Renilla ( n = 3, p

Techniques Used: Activity Assay, Transfection, Expressing, Luciferase

TET2 is required for Rosi-dependent gene regulation of PPARγ targets. Mature 3T3-L1 adipocytes were transduced with Tet2 -knockdown ( A-D ) or -overexpressor ( E-H ) lentiviruses. Two days after transduction, cells were treated with TNF (4 ng/ml), Rosi (1 μM), or both for 4 hours. Gene expression of a set of PPARγ target genes was determined by q-PCR ( n = 3, p
Figure Legend Snippet: TET2 is required for Rosi-dependent gene regulation of PPARγ targets. Mature 3T3-L1 adipocytes were transduced with Tet2 -knockdown ( A-D ) or -overexpressor ( E-H ) lentiviruses. Two days after transduction, cells were treated with TNF (4 ng/ml), Rosi (1 μM), or both for 4 hours. Gene expression of a set of PPARγ target genes was determined by q-PCR ( n = 3, p

Techniques Used: Transduction, Expressing, Polymerase Chain Reaction

22) Product Images from "Simvastatin and nanofibrous poly(L-lactic acid) scaffolds to promote the odontogenic potential of dental pulp cells in an inflammatory environment"

Article Title: Simvastatin and nanofibrous poly(L-lactic acid) scaffolds to promote the odontogenic potential of dental pulp cells in an inflammatory environment

Journal: Acta biomaterialia

doi: 10.1016/j.actbio.2017.12.037

Western blot results for pNFκBp65 (a) and PPARγ (b). The graphs show the quantification of protein expression by pixel intensity and is presented as fold increases compared to LPS−/SIM− group (negative control). The results
Figure Legend Snippet: Western blot results for pNFκBp65 (a) and PPARγ (b). The graphs show the quantification of protein expression by pixel intensity and is presented as fold increases compared to LPS−/SIM− group (negative control). The results

Techniques Used: Western Blot, Expressing, Negative Control

23) Product Images from "A Lox/CHOP‐10 crosstalk governs osteogenic and adipogenic cell fate by MSCs, et al. A Lox/CHOP‐10 crosstalk governs osteogenic and adipogenic cell fate by MSCs"

Article Title: A Lox/CHOP‐10 crosstalk governs osteogenic and adipogenic cell fate by MSCs, et al. A Lox/CHOP‐10 crosstalk governs osteogenic and adipogenic cell fate by MSCs

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13798

Lox inhibition together with BMP4 enhances bone formation in vivo. A, C3H10T1/2 stem cells transfected with Lox RNAi or negative RNAi in the presence of BMP4 were transplanted with HA‐TCP (hydroxyaptite‐tricalcium phosphate) subcutaneously into the armpit of immunocompromised mice for 4 wk. The transplants were harvested. Expression of osteogenic and adipogenic markers in transplants was evaluated by Q‐PCR (n = 4). B, BMP4 expression in the femur from WT and BMP4‐Tg mice. C, 3D micro‐CT images of cross sections (upper panel) and vertical sections (lower panel) of right femur extracted from BMP4‐Tg mice with PBS or BAPN treatment. Bars: upper panel, 200 μm; lower panel, 1 mm. D, Quantification of trabecular and cortical bone for the same femur as C. Results are shown as mean ± SEM. E, Western blotting of Lox, CHOP‐10, PPARγ, 422/aP2, Col1α1 and Ocn expression in the right femur extracted from BMP4‐Tg mice with PBS or BAPN treatment. * P
Figure Legend Snippet: Lox inhibition together with BMP4 enhances bone formation in vivo. A, C3H10T1/2 stem cells transfected with Lox RNAi or negative RNAi in the presence of BMP4 were transplanted with HA‐TCP (hydroxyaptite‐tricalcium phosphate) subcutaneously into the armpit of immunocompromised mice for 4 wk. The transplants were harvested. Expression of osteogenic and adipogenic markers in transplants was evaluated by Q‐PCR (n = 4). B, BMP4 expression in the femur from WT and BMP4‐Tg mice. C, 3D micro‐CT images of cross sections (upper panel) and vertical sections (lower panel) of right femur extracted from BMP4‐Tg mice with PBS or BAPN treatment. Bars: upper panel, 200 μm; lower panel, 1 mm. D, Quantification of trabecular and cortical bone for the same femur as C. Results are shown as mean ± SEM. E, Western blotting of Lox, CHOP‐10, PPARγ, 422/aP2, Col1α1 and Ocn expression in the right femur extracted from BMP4‐Tg mice with PBS or BAPN treatment. * P

Techniques Used: Inhibition, In Vivo, Transfection, Mouse Assay, Expressing, Polymerase Chain Reaction, Micro-CT, Western Blot

24) Product Images from "Inhibition of ASCT2 is essential in all-trans retinoic acid-induced reduction of adipogenesis in 3T3-L1 cells"

Article Title: Inhibition of ASCT2 is essential in all-trans retinoic acid-induced reduction of adipogenesis in 3T3-L1 cells

Journal: FEBS Open Bio

doi: 10.1016/j.fob.2015.06.012

Effects of ATRA on adipogenesis and Asct2 expression. (A) Lipid-droplets in the cultured 3T3-L1 cells detected with oil Red-O staining in insulin medium containing various concentrations of ATRA after 7 days. Each bar represents the mean ± SD. (B) Expression levels of asct2 during adipocyte differentiation in 3T3-L1 cells treated with 0.1% ethanol (EtOH) and 0.1 μM ATRA (day 5) as revealed by quantitative RT-PCR. (C) Immunoblot detection of Asct2 and PPARγ in 3T3-L1 cells treated with 0.1% EtOH and 1 μM ATRA (day 5). Gapdh was employed as an internal control. Graph shows the density of detected Asct2 normalized with Gapdh. The results are representative of three independent experiments. (D) Immunofluorescent staining of Asct2 in DIM-stimulated 3T3-L1 cells treated with 0.1% EtOH and 1 μM ATRA (day 5). Blue shows DAPI-stained nuclei. Red shows immunostained Asct2. Arrows indicate the presence of Asct2 on the cell surface of DIM-stimulated 3T3-L1 cells during adipogenesis.
Figure Legend Snippet: Effects of ATRA on adipogenesis and Asct2 expression. (A) Lipid-droplets in the cultured 3T3-L1 cells detected with oil Red-O staining in insulin medium containing various concentrations of ATRA after 7 days. Each bar represents the mean ± SD. (B) Expression levels of asct2 during adipocyte differentiation in 3T3-L1 cells treated with 0.1% ethanol (EtOH) and 0.1 μM ATRA (day 5) as revealed by quantitative RT-PCR. (C) Immunoblot detection of Asct2 and PPARγ in 3T3-L1 cells treated with 0.1% EtOH and 1 μM ATRA (day 5). Gapdh was employed as an internal control. Graph shows the density of detected Asct2 normalized with Gapdh. The results are representative of three independent experiments. (D) Immunofluorescent staining of Asct2 in DIM-stimulated 3T3-L1 cells treated with 0.1% EtOH and 1 μM ATRA (day 5). Blue shows DAPI-stained nuclei. Red shows immunostained Asct2. Arrows indicate the presence of Asct2 on the cell surface of DIM-stimulated 3T3-L1 cells during adipogenesis.

Techniques Used: Expressing, Cell Culture, Staining, Quantitative RT-PCR

25) Product Images from "The Expression of the Short Isoform of Thymic Stromal Lymphopoietin in the Colon Is Regulated by the Nuclear Receptor Peroxisome Proliferator Activated Receptor-Gamma and Is Impaired during Ulcerative Colitis"

Article Title: The Expression of the Short Isoform of Thymic Stromal Lymphopoietin in the Colon Is Regulated by the Nuclear Receptor Peroxisome Proliferator Activated Receptor-Gamma and Is Impaired during Ulcerative Colitis

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2017.01052

Activation of peroxisome proliferator activated receptor-gamma (PPARγ) in IEC cell lines induced TSLP expression. (A) qPCR analysis of TSLP gene expression in stimulated Caco-2 cells. Cells were stimulated for 24 h with each agonist. Results represent mean ± SEM (2 independent experiments in triplicate or sextuplicate, 9
Figure Legend Snippet: Activation of peroxisome proliferator activated receptor-gamma (PPARγ) in IEC cell lines induced TSLP expression. (A) qPCR analysis of TSLP gene expression in stimulated Caco-2 cells. Cells were stimulated for 24 h with each agonist. Results represent mean ± SEM (2 independent experiments in triplicate or sextuplicate, 9

Techniques Used: Activation Assay, Expressing, Real-time Polymerase Chain Reaction

sfTSLP expression is decreased in ulcerative colitis (UC) patients and correlates with peroxisome proliferator activated receptor-gamma (PPARγ) expression level. (A) Colonic mucosa were obtained from control subjects ( n = 22) or patients with UC (healthy mucosa, n = 9; injured mucosa, n = 21). Quantitative expression of PPARγ and sfTSLP mRNA were assessed by qPCR and normalized to GAPDH level. Transcript levels of sfTSLP in colonic mucosa from both controls and UC patients were correlated with PPARγ mRNA level. P -value was determined by the non-parametric Spearman test. (B) Colonic epithelial cells (CEC) were purified from resected colon specimens of control subjects ( n = 6) or untreated patients with UC ( n = 8). Quantitative expression of PPARγ and sfTSLP mRNA were assessed by qPCR and normalized to GAPDH level. Transcript levels of sfTSLP in CEC from both controls and UC patients were correlated with PPARγ mRNA level. P -value was determined by the non-parametric Spearman test. Horizontal bar indicates the mean value. ** P
Figure Legend Snippet: sfTSLP expression is decreased in ulcerative colitis (UC) patients and correlates with peroxisome proliferator activated receptor-gamma (PPARγ) expression level. (A) Colonic mucosa were obtained from control subjects ( n = 22) or patients with UC (healthy mucosa, n = 9; injured mucosa, n = 21). Quantitative expression of PPARγ and sfTSLP mRNA were assessed by qPCR and normalized to GAPDH level. Transcript levels of sfTSLP in colonic mucosa from both controls and UC patients were correlated with PPARγ mRNA level. P -value was determined by the non-parametric Spearman test. (B) Colonic epithelial cells (CEC) were purified from resected colon specimens of control subjects ( n = 6) or untreated patients with UC ( n = 8). Quantitative expression of PPARγ and sfTSLP mRNA were assessed by qPCR and normalized to GAPDH level. Transcript levels of sfTSLP in CEC from both controls and UC patients were correlated with PPARγ mRNA level. P -value was determined by the non-parametric Spearman test. Horizontal bar indicates the mean value. ** P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Capillary Electrochromatography, Purification

TSLP expression is strongly reduced in peroxisome proliferator activated receptor-gamma (PPARγ) knock-down Caco-2 cells. (A) Caco-2 colorectal cell line knock-down for PPARγ (ShPPARγ) expressed significantly fewer PPARγ transcripts and protein compared to ShLuc control cells. Quantitative expression of mRNA was assessed by quantitative PCR (qPCR). Results represent the mean values of sextuplicate ± SD of the fold change of PPARγ expression normalized to GAPDH level. The expression level measured in ShLuc cells (arbitrarily defined as one) was used as reference. Protein level was assessed by Western blot. The results represent a triplicate of the same clone of ShLuc and ShPPARγ Caco-2 cells, respectively. (B) Targeted approach in order to identify genes with different expression profile in PPARγ knock-down Caco-2 cells (ShPPARγ) compared to control cells (ShLuc)—Results of qPCR analysis. We selected various genes potentially expressed by intestinal epithelial cells and tested their expression in ShPPARγ/ShLuc Caco-2 cells. Results represent the mean ± SEM (2 independent experiments in triplicate) of the fold change of selected genes expression normalized to GAPDH level. The expression level measured in control cells was used as reference and defined as 1. (C) qPCR analysis of TSLP gene expression in PPARγ knock-down Caco-2 cells (ShPPARγ) compared to control cells (ShLuc). Results represent mean ± SEM (5 independent experiments in triplicate or sextuplicate, n = 18) of the fold change of TSLP gene expression normalized to GAPDH level. The expression level measured in control cells was used as reference and defined as 1. Caco-2 cells were stimulated for 24 h with GW9662. Results represent mean ± SEM (2 independent experiments in triplicate or sextuplicate, n = 9) of the fold change of TSLP gene expression normalized to GAPDH level. The expression level measured in control cells was used as reference and defined as 1. ** P
Figure Legend Snippet: TSLP expression is strongly reduced in peroxisome proliferator activated receptor-gamma (PPARγ) knock-down Caco-2 cells. (A) Caco-2 colorectal cell line knock-down for PPARγ (ShPPARγ) expressed significantly fewer PPARγ transcripts and protein compared to ShLuc control cells. Quantitative expression of mRNA was assessed by quantitative PCR (qPCR). Results represent the mean values of sextuplicate ± SD of the fold change of PPARγ expression normalized to GAPDH level. The expression level measured in ShLuc cells (arbitrarily defined as one) was used as reference. Protein level was assessed by Western blot. The results represent a triplicate of the same clone of ShLuc and ShPPARγ Caco-2 cells, respectively. (B) Targeted approach in order to identify genes with different expression profile in PPARγ knock-down Caco-2 cells (ShPPARγ) compared to control cells (ShLuc)—Results of qPCR analysis. We selected various genes potentially expressed by intestinal epithelial cells and tested their expression in ShPPARγ/ShLuc Caco-2 cells. Results represent the mean ± SEM (2 independent experiments in triplicate) of the fold change of selected genes expression normalized to GAPDH level. The expression level measured in control cells was used as reference and defined as 1. (C) qPCR analysis of TSLP gene expression in PPARγ knock-down Caco-2 cells (ShPPARγ) compared to control cells (ShLuc). Results represent mean ± SEM (5 independent experiments in triplicate or sextuplicate, n = 18) of the fold change of TSLP gene expression normalized to GAPDH level. The expression level measured in control cells was used as reference and defined as 1. Caco-2 cells were stimulated for 24 h with GW9662. Results represent mean ± SEM (2 independent experiments in triplicate or sextuplicate, n = 9) of the fold change of TSLP gene expression normalized to GAPDH level. The expression level measured in control cells was used as reference and defined as 1. ** P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Activation of peroxisome proliferator activated receptor-gamma (PPARγ) in IEC cell lines induced the expression of the short isoform of TSLP (sfTSLP). (A) Schematic representation of the structural organization of the TSLP gene with its two isoforms. Variant 1 represents the long form TSLP (lfTSLP) and variant 2 represents the short isoform (sfTSLP). The rectangles represent the exons, the gray parts corresponding to the untranslated regions and the black parts corresponding to the translated regions. The oligonucleotides used to amplify the TSLP gene are depicted. Oligonucleotides TSLP Fwd/TSLP Rev do not discriminate between both isoforms. Oligonucleotides TSLP1 Fwd/TSLP1 Rev specifically amplify lfTSLP and oligonucleotides TSLP2 Fwd/TSLP2 Rev specifically amplify sfTSLP. (B) qPCR analysis of sfTSLP gene expression in stimulated Caco-2 cells. Cells were stimulated for 24 h with each agonist. Results represent mean ± SEM (2 independent experiments in triplicate or sextuplicate, 9
Figure Legend Snippet: Activation of peroxisome proliferator activated receptor-gamma (PPARγ) in IEC cell lines induced the expression of the short isoform of TSLP (sfTSLP). (A) Schematic representation of the structural organization of the TSLP gene with its two isoforms. Variant 1 represents the long form TSLP (lfTSLP) and variant 2 represents the short isoform (sfTSLP). The rectangles represent the exons, the gray parts corresponding to the untranslated regions and the black parts corresponding to the translated regions. The oligonucleotides used to amplify the TSLP gene are depicted. Oligonucleotides TSLP Fwd/TSLP Rev do not discriminate between both isoforms. Oligonucleotides TSLP1 Fwd/TSLP1 Rev specifically amplify lfTSLP and oligonucleotides TSLP2 Fwd/TSLP2 Rev specifically amplify sfTSLP. (B) qPCR analysis of sfTSLP gene expression in stimulated Caco-2 cells. Cells were stimulated for 24 h with each agonist. Results represent mean ± SEM (2 independent experiments in triplicate or sextuplicate, 9

Techniques Used: Activation Assay, Expressing, Variant Assay, Real-time Polymerase Chain Reaction

Peroxisome proliferator activated receptor-gamma (PPARγ) binds functionally to the promoter of sfTSLP gene. (A) Schematic representation of the promoter of the sfTSLP gene (1,000 base pairs upstream of the transcription start site) showing the potential presence of PPAR response element predicted by in silico . (B) Picture showing PCR amplification of the 3a–4b fragment in ChIP assay from Caco-2 cells. “Control PCR” corresponds to the amplification of a genomic region unrelated to sfTSLP promoter and serves as a control of specificity for the ChIP analysis. (C) Schematic representation of the two reporter constructions used in transient transfection and reporter gene assays. (D,E) Luciferase gene reporter assays in Caco-2 cells transfected with PromTSLP2-Luc and PromTSLP2-LucΔ1 reporter constructs. Cells transfected with empty pGL4Luc plasmid were used as control. Results represent the mean ± SEM (3 independent experiments in triplicate or sextuplicate, 9
Figure Legend Snippet: Peroxisome proliferator activated receptor-gamma (PPARγ) binds functionally to the promoter of sfTSLP gene. (A) Schematic representation of the promoter of the sfTSLP gene (1,000 base pairs upstream of the transcription start site) showing the potential presence of PPAR response element predicted by in silico . (B) Picture showing PCR amplification of the 3a–4b fragment in ChIP assay from Caco-2 cells. “Control PCR” corresponds to the amplification of a genomic region unrelated to sfTSLP promoter and serves as a control of specificity for the ChIP analysis. (C) Schematic representation of the two reporter constructions used in transient transfection and reporter gene assays. (D,E) Luciferase gene reporter assays in Caco-2 cells transfected with PromTSLP2-Luc and PromTSLP2-LucΔ1 reporter constructs. Cells transfected with empty pGL4Luc plasmid were used as control. Results represent the mean ± SEM (3 independent experiments in triplicate or sextuplicate, 9

Techniques Used: In Silico, Polymerase Chain Reaction, Amplification, Chromatin Immunoprecipitation, Transfection, Luciferase, Construct, Plasmid Preparation

26) Product Images from "Schisandra chinensis Prevents Alcohol-Induced Fatty Liver Disease in Rats"

Article Title: Schisandra chinensis Prevents Alcohol-Induced Fatty Liver Disease in Rats

Journal: Journal of Medicinal Food

doi: 10.1089/jmf.2013.2849

Effect of SC on the expression of SREBP-1, AMPK, PPARα, and PPARγ in the liver. The representative Western blots from one of three independent experiments are shown. SC administration downregulated hepatic SREBP-1 and upregulated PPARα and phospho-AMPK expression in ethanol-fed rats. SREBP-1, sterol regulatory element-binding protein 1; AMPK, AMP-activated protein kinase; PPAR, peroxisome proliferator-activated receptor.
Figure Legend Snippet: Effect of SC on the expression of SREBP-1, AMPK, PPARα, and PPARγ in the liver. The representative Western blots from one of three independent experiments are shown. SC administration downregulated hepatic SREBP-1 and upregulated PPARα and phospho-AMPK expression in ethanol-fed rats. SREBP-1, sterol regulatory element-binding protein 1; AMPK, AMP-activated protein kinase; PPAR, peroxisome proliferator-activated receptor.

Techniques Used: Expressing, Western Blot, Binding Assay

27) Product Images from "PPARγ Regulates Pharmacological but not Physiological nor Pathological Osteoclast Formation"

Article Title: PPARγ Regulates Pharmacological but not Physiological nor Pathological Osteoclast Formation

Journal: Nature medicine

doi: 10.1038/nm.4208

PPARγ deficiency does not affect osteoclast differentiation in Vitro and in Vivo
Figure Legend Snippet: PPARγ deficiency does not affect osteoclast differentiation in Vitro and in Vivo

Techniques Used: In Vitro, In Vivo

28) Product Images from "Roles of PPAR?/NF-?B Signaling Pathway in the Pathogenesis of Intrahepatic Cholestasis of Pregnancy"

Article Title: Roles of PPAR?/NF-?B Signaling Pathway in the Pathogenesis of Intrahepatic Cholestasis of Pregnancy

Journal: PLoS ONE

doi: 10.1371/journal.pone.0087343

Expression of PPARγ and NF-κB protein in cultured HTR-8/SVneo cell. (A) Western blotting analysis of placental PPARγ and NF-κB protein expression in control, mild ICP and sever ICP group. (B) Graphical summary of data on the expression of PPARγ mRNA. (C) Graphical summary of data on the expression of NF-κB protein. The data are expressed as the mean ± S.D., ## p
Figure Legend Snippet: Expression of PPARγ and NF-κB protein in cultured HTR-8/SVneo cell. (A) Western blotting analysis of placental PPARγ and NF-κB protein expression in control, mild ICP and sever ICP group. (B) Graphical summary of data on the expression of PPARγ mRNA. (C) Graphical summary of data on the expression of NF-κB protein. The data are expressed as the mean ± S.D., ## p

Techniques Used: Expressing, Cell Culture, Western Blot

Expression of PPARγ and NF-κB protein in placentas from control group and ICP groups. (A) Western blotting analysis of placental PPARγ and NF-κB protein expression in control, mild ICP and sever ICP groups. (B) Graphical summary of data on the expression of PPARγ protein. (C) Graphical summary of data on the expression of NF-κB protein. The data are expressed as the mean ± S.D., ** p
Figure Legend Snippet: Expression of PPARγ and NF-κB protein in placentas from control group and ICP groups. (A) Western blotting analysis of placental PPARγ and NF-κB protein expression in control, mild ICP and sever ICP groups. (B) Graphical summary of data on the expression of PPARγ protein. (C) Graphical summary of data on the expression of NF-κB protein. The data are expressed as the mean ± S.D., ** p

Techniques Used: Expressing, Western Blot

Expression of PPARγ andNF-κB mRNA in cultured HTR-8/SVneo cell. (A) RT-PCR analysis of placental PPARγ and NF-κB mRNA expression in control, mild ICP and sever ICP group. (B) Graphical summary of data on the expression of PPARγ mRNA. (C) Graphical summary of data on the expression of NF-κB mRNA. The data are expressed as the mean ± S.D., ## p
Figure Legend Snippet: Expression of PPARγ andNF-κB mRNA in cultured HTR-8/SVneo cell. (A) RT-PCR analysis of placental PPARγ and NF-κB mRNA expression in control, mild ICP and sever ICP group. (B) Graphical summary of data on the expression of PPARγ mRNA. (C) Graphical summary of data on the expression of NF-κB mRNA. The data are expressed as the mean ± S.D., ## p

Techniques Used: Expressing, Cell Culture, Reverse Transcription Polymerase Chain Reaction

PPARγ and NF-κB staining were found in the membrane and cytoplasm of placental trophoblast cell. ×400 (A–C) PPARγ protein expressed in the placenta of control patients, mild ICP patients and severe ICP patients. (D–F) NF-κB protein expressed in the placenta of control patients mild ICP patients and severe ICP patients. PPARγ and NF-κB proteins expression were significantly different in control group and ICP groups.
Figure Legend Snippet: PPARγ and NF-κB staining were found in the membrane and cytoplasm of placental trophoblast cell. ×400 (A–C) PPARγ protein expressed in the placenta of control patients, mild ICP patients and severe ICP patients. (D–F) NF-κB protein expressed in the placenta of control patients mild ICP patients and severe ICP patients. PPARγ and NF-κB proteins expression were significantly different in control group and ICP groups.

Techniques Used: Staining, Expressing

Expression of PPARγ and NF-κB mRNA in placentas from control group and ICP groups. (A) RT-PCR analysis of placental PPARγ and NF-κB mRNAexpression in control, mild ICP and sever ICP groups. (B) Graphical summary of data on the expression of PPARγ mRNA. (C) Graphical summary of data on the expression of NF-κB mRNA. The data are expressed as the mean ± S.D., ** p
Figure Legend Snippet: Expression of PPARγ and NF-κB mRNA in placentas from control group and ICP groups. (A) RT-PCR analysis of placental PPARγ and NF-κB mRNAexpression in control, mild ICP and sever ICP groups. (B) Graphical summary of data on the expression of PPARγ mRNA. (C) Graphical summary of data on the expression of NF-κB mRNA. The data are expressed as the mean ± S.D., ** p

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

29) Product Images from "Cardiac protective effects of irbesartan via the PPAR-gamma signaling pathway in angiotensin-converting enzyme 2-deficient mice"

Article Title: Cardiac protective effects of irbesartan via the PPAR-gamma signaling pathway in angiotensin-converting enzyme 2-deficient mice

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-11-229

Cardiac protein levels of PPARα, PPARδ and PPARγ in mice. Representative Western blot (A) exhibited cardiac protein expression (B) of PPARα, PPARδ, and PPARγ in mice. α-tubulin was used as endogenous control. Irb = irbesartan, PPAR, peroxisome proliferator-activated receptor. n = 5 for each group. ** P
Figure Legend Snippet: Cardiac protein levels of PPARα, PPARδ and PPARγ in mice. Representative Western blot (A) exhibited cardiac protein expression (B) of PPARα, PPARδ, and PPARγ in mice. α-tubulin was used as endogenous control. Irb = irbesartan, PPAR, peroxisome proliferator-activated receptor. n = 5 for each group. ** P

Techniques Used: Mouse Assay, Western Blot, Expressing

30) Product Images from "ENHANCING ADULT NERVE REGENERATION THROUGH THE KNOCKDOWN OF RETINOBLASTOMA PROTEIN"

Article Title: ENHANCING ADULT NERVE REGENERATION THROUGH THE KNOCKDOWN OF RETINOBLASTOMA PROTEIN

Journal: Nature communications

doi: 10.1038/ncomms4670

Neurite plasticity following Rb1 knockdown is blocked by a PPARγ antagonist Dissociated uninjured (non-preconditioned) adult sensory neurons exposed to one of two differing Rb1 siRNA constructs showed enhanced growth. Bars indicate control neurite outgrowth, Rb1 siRNA outgrowth and Rb1 siRNA outgrowth with GW9662 at 0.1 or 1.0 μM. Rb1 siRNA is associated with rises in total neurite outgrowth (a) [*p
Figure Legend Snippet: Neurite plasticity following Rb1 knockdown is blocked by a PPARγ antagonist Dissociated uninjured (non-preconditioned) adult sensory neurons exposed to one of two differing Rb1 siRNA constructs showed enhanced growth. Bars indicate control neurite outgrowth, Rb1 siRNA outgrowth and Rb1 siRNA outgrowth with GW9662 at 0.1 or 1.0 μM. Rb1 siRNA is associated with rises in total neurite outgrowth (a) [*p

Techniques Used: Construct

Knockdown of Rb1 increases the expression of PPARγ Rb1 siRNA is associated with knockdown of Rb1 protein in dissociated adult sensory neurons (a) and a rise in PPARγ expression (b) without changes in p-Akt, Pak1 or CDK5 (c,d) . Quantitation of western blot data showing declines in Rb (e) [ *p
Figure Legend Snippet: Knockdown of Rb1 increases the expression of PPARγ Rb1 siRNA is associated with knockdown of Rb1 protein in dissociated adult sensory neurons (a) and a rise in PPARγ expression (b) without changes in p-Akt, Pak1 or CDK5 (c,d) . Quantitation of western blot data showing declines in Rb (e) [ *p

Techniques Used: Expressing, Quantitation Assay, Western Blot

31) Product Images from "Cyclic Stretch Facilitates Myogenesis in C2C12 Myoblasts and Rescues Thiazolidinedione-Inhibited Myotube Formation"

Article Title: Cyclic Stretch Facilitates Myogenesis in C2C12 Myoblasts and Rescues Thiazolidinedione-Inhibited Myotube Formation

Journal: Frontiers in Bioengineering and Biotechnology

doi: 10.3389/fbioe.2016.00027

PPARγ protein expression was decreased during myogenesis (A) . The transcriptional activity of PPARs was decreased as measured by the promoter luciferase reporter construct of the PPAR-responsive element (PPRE) (B) . *Significant difference compared with GM ( p
Figure Legend Snippet: PPARγ protein expression was decreased during myogenesis (A) . The transcriptional activity of PPARs was decreased as measured by the promoter luciferase reporter construct of the PPAR-responsive element (PPRE) (B) . *Significant difference compared with GM ( p

Techniques Used: Expressing, Activity Assay, Luciferase, Construct

After applying the cyclic stretch for 1 h, α-SMA protein expression was increased with TZD treatment in MM (A) . The number below each lane indicates the quantified fold change with normalized to static condition without TZD and its individual β-actin. The mechanical stretch also inhibited the TZD-induced PPRE promoter activity (B) . The decreased PPARγ protein expression can be reversed by adding the protease inhibitor MG-132 during the cyclic stretch (C) . *Significant difference compared with GM without drugs ( p
Figure Legend Snippet: After applying the cyclic stretch for 1 h, α-SMA protein expression was increased with TZD treatment in MM (A) . The number below each lane indicates the quantified fold change with normalized to static condition without TZD and its individual β-actin. The mechanical stretch also inhibited the TZD-induced PPRE promoter activity (B) . The decreased PPARγ protein expression can be reversed by adding the protease inhibitor MG-132 during the cyclic stretch (C) . *Significant difference compared with GM without drugs ( p

Techniques Used: Expressing, Activity Assay, Protease Inhibitor

Application of cyclic stretch (10% strain at 1 Hz for 1 h) in MM with further induced myogenesis as indicated by increased α-SMA expression (A) . The phosphorylation of JNK was induced by applying stretch in MM, whereas ERK phosphorylation was observed when stretch was applied in GM. A decrease in PPARγ protein expression was also found during stretch-induced myogenesis in MM (B) . The number below each lane indicates the quantified fold change with normalized to GM static condition and its individual β-actin. A further decrease of PPRE promoter activity was detected when applying cyclic stretch (C) . *Significant difference compared with GM ( p
Figure Legend Snippet: Application of cyclic stretch (10% strain at 1 Hz for 1 h) in MM with further induced myogenesis as indicated by increased α-SMA expression (A) . The phosphorylation of JNK was induced by applying stretch in MM, whereas ERK phosphorylation was observed when stretch was applied in GM. A decrease in PPARγ protein expression was also found during stretch-induced myogenesis in MM (B) . The number below each lane indicates the quantified fold change with normalized to GM static condition and its individual β-actin. A further decrease of PPRE promoter activity was detected when applying cyclic stretch (C) . *Significant difference compared with GM ( p

Techniques Used: Expressing, Activity Assay

The addition of TZD (PPARγ agonist) abolished the MM-induced myotube formation, but GW9662 (GW, PPARγ antagonist) increased the number and length of myotubes (A) . The effect of PPARγ on myogenesis was confirmed by MHC protein expression, which was decreased with TZD and increased with GW treatment (B) . The administration of TZD significantly increased PPRE promoter activity (C) . *Significant difference compared with GM without drugs (Ct) ( p
Figure Legend Snippet: The addition of TZD (PPARγ agonist) abolished the MM-induced myotube formation, but GW9662 (GW, PPARγ antagonist) increased the number and length of myotubes (A) . The effect of PPARγ on myogenesis was confirmed by MHC protein expression, which was decreased with TZD and increased with GW treatment (B) . The administration of TZD significantly increased PPRE promoter activity (C) . *Significant difference compared with GM without drugs (Ct) ( p

Techniques Used: Expressing, Activity Assay

32) Product Images from "LRP1 integrates murine macrophage cholesterol homeostasis and inflammatory responses in atherosclerosis"

Article Title: LRP1 integrates murine macrophage cholesterol homeostasis and inflammatory responses in atherosclerosis

Journal: eLife

doi: 10.7554/eLife.29292

Role of LRP1 NPxY phosphorylation in macrophage lipid accumulation and atherogenesis. ( A ) Tyrosine phosphorylation of LRP1 NPxY motifs (as indicated by asterisks), specifically the distal NPxY motif, is necessary for interaction with Shc1, which in turn activates PI3K/Akt signaling. Phospho-Akt modulates PPARγ/LXR driven Abca1 gene transcription and is necessary for the full effect of ox-LDL and PPARγ/LXR induced expression of Abca1 to mediate cholesterol efflux. Inhibition of PI3K/Akt (using LY294002 or Akt inhibitor) results in only partial induction of Abca 1 in response to ox-LDL and PPARγ/LXR agonists (rosiglitazone, T0901317, LXR623). ( B ) LRP1 integrates inflammatory signals and cholesterol homeostasis in macrophages. Inflammatory signals are initiated by the interaction between oxLDL and CD36/TLR4-6 in lipid rafts. In the setting of inflammation or cholesterol loading, increased proximity of LRP1 with activated SFKs in lipid rafts favors LRP1 tyrosine phosphorylation. This, in turn, activates a Shc1/PI3K/Akt/Pparγ/Lxrα axis that promotes Abca1 expression and cellular cholesterol export, which then leads to reduction of lipid raft cholesterol content and dissociation of LRP1 from the lipid raft, thus creating a negative feedback loop. LRP1 can also be cleaved by γ–secretase, thereby releasing the intracellular domain (ICD) which translocates to the nucleus where it suppresses inflammatory gene expression ( Zurhove et al., 2008 ). Whether the LRP1 ICD, in a phosphorylated or unphosphorylated state, can also alter ABCA1 expression is currently unknown. CD36, cluster of differentiation 36; TLR, toll like receptor; MyD88, myeloid differentiation primary response protein 88; TRIF, TIR domain-containing adaptor protein inducing IFNβ; SFK, SRC family kinase. ( C ) LRP1 regulates clearance of apoptotic cells (AC) through efferocytosis. LRP1 coordinates with other receptors, such AXL and MerTK, and adaptors such as RanBP9 and GULP, to recognize and engulf apoptotic cells. Engulfment of apoptotic cells is a critical step in the activation of LXR-responsive genes, including Abca1 and Mertk . PS, protein S; CRT, cell surface calreticulin; GAS6, growth arrest-specific 6; AXL, receptor tyrosine kinase; MerTK, Proto-oncogene tyrosine-protein kinase; RANBP9, RAN binding protein 9; GULP, engulfment adaptor PTB domain containing one.
Figure Legend Snippet: Role of LRP1 NPxY phosphorylation in macrophage lipid accumulation and atherogenesis. ( A ) Tyrosine phosphorylation of LRP1 NPxY motifs (as indicated by asterisks), specifically the distal NPxY motif, is necessary for interaction with Shc1, which in turn activates PI3K/Akt signaling. Phospho-Akt modulates PPARγ/LXR driven Abca1 gene transcription and is necessary for the full effect of ox-LDL and PPARγ/LXR induced expression of Abca1 to mediate cholesterol efflux. Inhibition of PI3K/Akt (using LY294002 or Akt inhibitor) results in only partial induction of Abca 1 in response to ox-LDL and PPARγ/LXR agonists (rosiglitazone, T0901317, LXR623). ( B ) LRP1 integrates inflammatory signals and cholesterol homeostasis in macrophages. Inflammatory signals are initiated by the interaction between oxLDL and CD36/TLR4-6 in lipid rafts. In the setting of inflammation or cholesterol loading, increased proximity of LRP1 with activated SFKs in lipid rafts favors LRP1 tyrosine phosphorylation. This, in turn, activates a Shc1/PI3K/Akt/Pparγ/Lxrα axis that promotes Abca1 expression and cellular cholesterol export, which then leads to reduction of lipid raft cholesterol content and dissociation of LRP1 from the lipid raft, thus creating a negative feedback loop. LRP1 can also be cleaved by γ–secretase, thereby releasing the intracellular domain (ICD) which translocates to the nucleus where it suppresses inflammatory gene expression ( Zurhove et al., 2008 ). Whether the LRP1 ICD, in a phosphorylated or unphosphorylated state, can also alter ABCA1 expression is currently unknown. CD36, cluster of differentiation 36; TLR, toll like receptor; MyD88, myeloid differentiation primary response protein 88; TRIF, TIR domain-containing adaptor protein inducing IFNβ; SFK, SRC family kinase. ( C ) LRP1 regulates clearance of apoptotic cells (AC) through efferocytosis. LRP1 coordinates with other receptors, such AXL and MerTK, and adaptors such as RanBP9 and GULP, to recognize and engulf apoptotic cells. Engulfment of apoptotic cells is a critical step in the activation of LXR-responsive genes, including Abca1 and Mertk . PS, protein S; CRT, cell surface calreticulin; GAS6, growth arrest-specific 6; AXL, receptor tyrosine kinase; MerTK, Proto-oncogene tyrosine-protein kinase; RANBP9, RAN binding protein 9; GULP, engulfment adaptor PTB domain containing one.

Techniques Used: Expressing, Inhibition, Activation Assay, Binding Assay

Lrp1 Y63F impairs Abca1 induction through inhibiting the PPARγ/LXR pathway. ( A ) Real-time PCR analysis of Abca1, Abca7, Pparg, Nr1h3 and Nr2h2 expression in macrophages treated with or without ox-LDL. ( B–E ) Isolated peritoneal macrophages from Lrp1 Y63F ;Ldlr −/− and Ldlr −/− were pre-incubated for 1 hr with ( B ) T0070907 (PPARγ antagonist, 10 μM), ( C ) Rosiglitizone (PPARγ agonist) at indicated concentrations in μM, ( D ) T0901317 (nonspecific LXR agonist) at indicated concentrations in μM, ( E ) LXR623 (LXRβ full agonist and LXRα partial agonist) at indicated concentrations in μM, followed by the treatment with or without oxLDL for an additional 24 hr. Abca1 expression was assessed by immunoblot analysis (left). Protein expression was quantified and analyzed (right). Data are mean ±SEM from three independent experiments. *p
Figure Legend Snippet: Lrp1 Y63F impairs Abca1 induction through inhibiting the PPARγ/LXR pathway. ( A ) Real-time PCR analysis of Abca1, Abca7, Pparg, Nr1h3 and Nr2h2 expression in macrophages treated with or without ox-LDL. ( B–E ) Isolated peritoneal macrophages from Lrp1 Y63F ;Ldlr −/− and Ldlr −/− were pre-incubated for 1 hr with ( B ) T0070907 (PPARγ antagonist, 10 μM), ( C ) Rosiglitizone (PPARγ agonist) at indicated concentrations in μM, ( D ) T0901317 (nonspecific LXR agonist) at indicated concentrations in μM, ( E ) LXR623 (LXRβ full agonist and LXRα partial agonist) at indicated concentrations in μM, followed by the treatment with or without oxLDL for an additional 24 hr. Abca1 expression was assessed by immunoblot analysis (left). Protein expression was quantified and analyzed (right). Data are mean ±SEM from three independent experiments. *p

Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Isolation, Incubation

33) Product Images from "The effect of dehydroleucodine in adipocyte differentiation."

Article Title: The effect of dehydroleucodine in adipocyte differentiation.

Journal: European Journal of Pharmacology

doi: 10.1016/j.ejphar.2011.09.033

Dehydroleucodine attenuated the expression of PPARγ during 3T3-L1 preadipocyte differentiation 3T3-L1 preadipocytes were induced to differentiate by induction media into adipocytes in the absence (insert: –DhL, control) or in the presence (insert: +DhL) of 8 μM DhL. Total protein extracts were prepared at day 9 from each sample. The proteins were subset to 12% SDS-PAGE electrophoresis, blotted to a nitrocellulose membrane, and probed with anti-bodies specific to (A) PPARγ, C-EBPα and tubulin, (B) P-Erk1/2, T-Erk1/2, (C) P-Akt1, T-Akt1, and (D) P-AMPKα and T-AMPKα respectively. Relative levels of proteins were determined by densitometry as describe in Material and Methods. Data represent the mean ± S.E.M. of three independent experiments. *P
Figure Legend Snippet: Dehydroleucodine attenuated the expression of PPARγ during 3T3-L1 preadipocyte differentiation 3T3-L1 preadipocytes were induced to differentiate by induction media into adipocytes in the absence (insert: –DhL, control) or in the presence (insert: +DhL) of 8 μM DhL. Total protein extracts were prepared at day 9 from each sample. The proteins were subset to 12% SDS-PAGE electrophoresis, blotted to a nitrocellulose membrane, and probed with anti-bodies specific to (A) PPARγ, C-EBPα and tubulin, (B) P-Erk1/2, T-Erk1/2, (C) P-Akt1, T-Akt1, and (D) P-AMPKα and T-AMPKα respectively. Relative levels of proteins were determined by densitometry as describe in Material and Methods. Data represent the mean ± S.E.M. of three independent experiments. *P

Techniques Used: Expressing, SDS Page, Electrophoresis

34) Product Images from "Isobavachalcone from Angelica keiskei Inhibits Adipogenesis and Prevents Lipid Accumulation"

Article Title: Isobavachalcone from Angelica keiskei Inhibits Adipogenesis and Prevents Lipid Accumulation

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19061693

Effect of IBC on autophagic flux during adipocyte differentiation. ( A ) 3T3-L1 adipocytes were differentiated in the presence or absence of 40 μM of IBC. Differentiating adipocytes were harvested at the indicated period and lysed for Western blotting analysis to determine levels of LC3B and SQSTM1/p62; ( B ) Preadipocytes were transfected with GFP-LC3 plasmid and differentiated with MDI in the presence of IBC or CQ (10 μM) during D0–D2, followed by additional treatment with differentiation medium for 2 days (D4). Cells were fixed and stained with DAPI (nuclei, blue-colored). Intracellular GFP-LC3 puncta (green-colored) were visualized with a confocal laser microscope. Scale bar = 10 μm; ( C ) Differentiating adipocytes supplemented with IBC or CQ during D0–D2 were harvested and protein levels of PPARγ and SQSTM1/p62 were analyzed on D8. Scale bar = 100 μm; and ( D ) On D2, differentiating adipocytes supplemented with IBC or CQ were harvested and subjected to qPCR to analyze gene expression levels of BECN1 , Atg5 , and Atg7 . Data are presented as means ± SD of triplicate experiments. * p
Figure Legend Snippet: Effect of IBC on autophagic flux during adipocyte differentiation. ( A ) 3T3-L1 adipocytes were differentiated in the presence or absence of 40 μM of IBC. Differentiating adipocytes were harvested at the indicated period and lysed for Western blotting analysis to determine levels of LC3B and SQSTM1/p62; ( B ) Preadipocytes were transfected with GFP-LC3 plasmid and differentiated with MDI in the presence of IBC or CQ (10 μM) during D0–D2, followed by additional treatment with differentiation medium for 2 days (D4). Cells were fixed and stained with DAPI (nuclei, blue-colored). Intracellular GFP-LC3 puncta (green-colored) were visualized with a confocal laser microscope. Scale bar = 10 μm; ( C ) Differentiating adipocytes supplemented with IBC or CQ during D0–D2 were harvested and protein levels of PPARγ and SQSTM1/p62 were analyzed on D8. Scale bar = 100 μm; and ( D ) On D2, differentiating adipocytes supplemented with IBC or CQ were harvested and subjected to qPCR to analyze gene expression levels of BECN1 , Atg5 , and Atg7 . Data are presented as means ± SD of triplicate experiments. * p

Techniques Used: Western Blot, Transfection, Plasmid Preparation, Staining, Microscopy, Real-time Polymerase Chain Reaction, Expressing

35) Product Images from "A Lox/CHOP‐10 crosstalk governs osteogenic and adipogenic cell fate by MSCs, et al. A Lox/CHOP‐10 crosstalk governs osteogenic and adipogenic cell fate by MSCs"

Article Title: A Lox/CHOP‐10 crosstalk governs osteogenic and adipogenic cell fate by MSCs, et al. A Lox/CHOP‐10 crosstalk governs osteogenic and adipogenic cell fate by MSCs

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13798

Lox inhibition together with BMP4 enhances bone formation in vivo. A, C3H10T1/2 stem cells transfected with Lox RNAi or negative RNAi in the presence of BMP4 were transplanted with HA‐TCP (hydroxyaptite‐tricalcium phosphate) subcutaneously into the armpit of immunocompromised mice for 4 wk. The transplants were harvested. Expression of osteogenic and adipogenic markers in transplants was evaluated by Q‐PCR (n = 4). B, BMP4 expression in the femur from WT and BMP4‐Tg mice. C, 3D micro‐CT images of cross sections (upper panel) and vertical sections (lower panel) of right femur extracted from BMP4‐Tg mice with PBS or BAPN treatment. Bars: upper panel, 200 μm; lower panel, 1 mm. D, Quantification of trabecular and cortical bone for the same femur as C. Results are shown as mean ± SEM. E, Western blotting of Lox, CHOP‐10, PPARγ, 422/aP2, Col1α1 and Ocn expression in the right femur extracted from BMP4‐Tg mice with PBS or BAPN treatment. * P
Figure Legend Snippet: Lox inhibition together with BMP4 enhances bone formation in vivo. A, C3H10T1/2 stem cells transfected with Lox RNAi or negative RNAi in the presence of BMP4 were transplanted with HA‐TCP (hydroxyaptite‐tricalcium phosphate) subcutaneously into the armpit of immunocompromised mice for 4 wk. The transplants were harvested. Expression of osteogenic and adipogenic markers in transplants was evaluated by Q‐PCR (n = 4). B, BMP4 expression in the femur from WT and BMP4‐Tg mice. C, 3D micro‐CT images of cross sections (upper panel) and vertical sections (lower panel) of right femur extracted from BMP4‐Tg mice with PBS or BAPN treatment. Bars: upper panel, 200 μm; lower panel, 1 mm. D, Quantification of trabecular and cortical bone for the same femur as C. Results are shown as mean ± SEM. E, Western blotting of Lox, CHOP‐10, PPARγ, 422/aP2, Col1α1 and Ocn expression in the right femur extracted from BMP4‐Tg mice with PBS or BAPN treatment. * P

Techniques Used: Inhibition, In Vivo, Transfection, Mouse Assay, Expressing, Polymerase Chain Reaction, Micro-CT, Western Blot

High‐fat diet mice displayed bone loss accompanied with Lox induction. A, 3D micro‐CT images of cross sections (upper panel) and vertical sections (lower panel) of right femur extracted from NCD mice or HFD mice. Bars: upper panel, 200 μm; lower panel, 1 mm. B, Quantification of trabecular and cortical bone for the same femur as shown in Figure 1 A. C, Q‐PCR analysis of Lox, PPARγ, 422/aP2, Col1α1 and Ocn expression in femurs of NCD (n = 5) and HFD (n = 5) mice. D, Western blotting of Lox, PPARγ, 422/aP2, Col1α1 and Ocn expression in femurs of NCD (n = 3) and HFD (n = 3) mice. NCD, normal chow diet; HFD, high‐fat diet. E, Graphical abstract. * P
Figure Legend Snippet: High‐fat diet mice displayed bone loss accompanied with Lox induction. A, 3D micro‐CT images of cross sections (upper panel) and vertical sections (lower panel) of right femur extracted from NCD mice or HFD mice. Bars: upper panel, 200 μm; lower panel, 1 mm. B, Quantification of trabecular and cortical bone for the same femur as shown in Figure 1 A. C, Q‐PCR analysis of Lox, PPARγ, 422/aP2, Col1α1 and Ocn expression in femurs of NCD (n = 5) and HFD (n = 5) mice. D, Western blotting of Lox, PPARγ, 422/aP2, Col1α1 and Ocn expression in femurs of NCD (n = 3) and HFD (n = 3) mice. NCD, normal chow diet; HFD, high‐fat diet. E, Graphical abstract. * P

Techniques Used: Mouse Assay, Micro-CT, Polymerase Chain Reaction, Expressing, Western Blot

36) Product Images from "Differential transcription factor expression by human epithelial cells of buccal and urothelial derivation"

Article Title: Differential transcription factor expression by human epithelial cells of buccal and urothelial derivation

Journal: Experimental Cell Research

doi: 10.1016/j.yexcr.2018.05.031

Evaluation of PPARγ, FOXA1 and GATA3 expression in PPARG1 overexpressing and control (empty) NHB cells. PPARG1 overexpressing and control (empty vector) NHB cell cultures were exposed to the TZ/PD protocol for 72 h. Experiments performed on between 2 and 4 independent NHB cell lines (as stated below), with representative results shown. (A) PPARγ1, FOXA1 and GATA3 protein expression assessed by western blotting. ACTB expression was included as an internal loading control. Protein lysates from cell lines known to express the proteins of interest were included as positive controls for each antibody (CTRL). Experiments performed on n = 3 independent NHB cell lines. (B) Densitometry analysis of FOXA1 protein expression shown relative to control (Empty - DMSO) NHB cells. Data is shown as the mean of n = 4 independent transduced NHB cell lines. All values were normalised to the ACTB expression. Statistical analysis was performed using a one-way ANOVA test, but no statistical significance was found (P > 0.05). Error bars represent standard deviation. (C) Immunofluorescence microscopy evaluating PPARγ, FOXA1 and GATA3 protein localisation in PPARG1 overexpressing and control (empty vector) NHB cells following the TZ/PD protocol at 72 h. Experiments were performed on n = 3 independent transduced NHB cell lines. IF images for a single NHB cell line are shown. Nuclear localisation was observed with PPARγ (n = 3/3), FOXA1 (n = 3/3), and GATA3 (n = 2/3). NHU cells (non-transduced) treated with the TZ/PD protocol are shown for comparison at the same time point. Scale bar ≡ 50 µm.
Figure Legend Snippet: Evaluation of PPARγ, FOXA1 and GATA3 expression in PPARG1 overexpressing and control (empty) NHB cells. PPARG1 overexpressing and control (empty vector) NHB cell cultures were exposed to the TZ/PD protocol for 72 h. Experiments performed on between 2 and 4 independent NHB cell lines (as stated below), with representative results shown. (A) PPARγ1, FOXA1 and GATA3 protein expression assessed by western blotting. ACTB expression was included as an internal loading control. Protein lysates from cell lines known to express the proteins of interest were included as positive controls for each antibody (CTRL). Experiments performed on n = 3 independent NHB cell lines. (B) Densitometry analysis of FOXA1 protein expression shown relative to control (Empty - DMSO) NHB cells. Data is shown as the mean of n = 4 independent transduced NHB cell lines. All values were normalised to the ACTB expression. Statistical analysis was performed using a one-way ANOVA test, but no statistical significance was found (P > 0.05). Error bars represent standard deviation. (C) Immunofluorescence microscopy evaluating PPARγ, FOXA1 and GATA3 protein localisation in PPARG1 overexpressing and control (empty vector) NHB cells following the TZ/PD protocol at 72 h. Experiments were performed on n = 3 independent transduced NHB cell lines. IF images for a single NHB cell line are shown. Nuclear localisation was observed with PPARγ (n = 3/3), FOXA1 (n = 3/3), and GATA3 (n = 2/3). NHU cells (non-transduced) treated with the TZ/PD protocol are shown for comparison at the same time point. Scale bar ≡ 50 µm.

Techniques Used: Expressing, Plasmid Preparation, Western Blot, Standard Deviation, Immunofluorescence, Microscopy

GATA3 overexpression in NHB cells. (A) GATA3 overexpressing and control (empty vector) NHB cell cultures following exposure to the PPARγ-activating TZ/PD protocol for 72 h. Western blotting of whole protein lysates was performed to assess protein expression of GATA3, FOXA1 and PPARy1. NHU cells (non-transduced) and treated with the TZ/PD protocol for 72 h are shown for comparison. (B) GATA3 overexpressing and control (empty vector) NHB cells at 72 h post TZ/PD protocol. GATA3, FOXA1 and PPARy protein expression assessed by indirect immunofluorescence microscopy. NHU cells (non-transduced; 72 h TZ/PD protocol) were included as positive controls for comparison. Scale bar ≡ 50 µm. (C) GATA3 overexpressing and control (empty vector) NHB cells were induced to form cell sheets using 5% ABS and 2 mM calcium for up to 7 days. Expression of the tight junction-associated proteins, claudin 3, 4, 5 and 7, assessed by western blotting. ACTB was included as a loading control. NHU cells (non-transduced) exposed to the same protocol were used as a positive control for comparison. Experiments were performed on n = 2 independent NHB donor cell lines and representative results shown.
Figure Legend Snippet: GATA3 overexpression in NHB cells. (A) GATA3 overexpressing and control (empty vector) NHB cell cultures following exposure to the PPARγ-activating TZ/PD protocol for 72 h. Western blotting of whole protein lysates was performed to assess protein expression of GATA3, FOXA1 and PPARy1. NHU cells (non-transduced) and treated with the TZ/PD protocol for 72 h are shown for comparison. (B) GATA3 overexpressing and control (empty vector) NHB cells at 72 h post TZ/PD protocol. GATA3, FOXA1 and PPARy protein expression assessed by indirect immunofluorescence microscopy. NHU cells (non-transduced; 72 h TZ/PD protocol) were included as positive controls for comparison. Scale bar ≡ 50 µm. (C) GATA3 overexpressing and control (empty vector) NHB cells were induced to form cell sheets using 5% ABS and 2 mM calcium for up to 7 days. Expression of the tight junction-associated proteins, claudin 3, 4, 5 and 7, assessed by western blotting. ACTB was included as a loading control. NHU cells (non-transduced) exposed to the same protocol were used as a positive control for comparison. Experiments were performed on n = 2 independent NHB donor cell lines and representative results shown.

Techniques Used: Over Expression, Plasmid Preparation, Western Blot, Expressing, Immunofluorescence, Microscopy, Positive Control

Comparison of uroplakin and urothelium differentiation-associated transcription factor gene expression by NHB and NHU cell cultures. Employing protocols developed to differentiate NHU cells by PPARγ activation, cell cultures of NHB or NHU cells were exposed to 1 µM troglitazone and 1 µM PD153035 (TZ/PD) for 24 h, maintained in 1 µM PD153035 and harvested at 12, 24, 48 and/or 72 h. Control cultures were exposed to vehicle (0.1% DMSO) alone. (A) RTqPCR for three independent NHB cell lines (represented by different symbols), versus a single NHU cell line for comparison of uroplakin (UPK1A, UPK1B, UPK2, UPK3A and UPK3B) mRNA expression at the 72 h time-point. All data has been normalised to GAPDH expression and is presented relative to the DMSO-treated NHB cells for each gene except UPK3A, where the data is shown relative to the DMSO treated NHU cells due to absent UPK3A gene expression by NHB cells. BLOD = Below Limit of Detection. Statistical analysis was performed using a two-tailed, paired t -test to determine whether TZ/PD resulted in any significant change in gene expression in NHB cells. *represents P ≤ 0.05, ** represents P ≤ 0.01. Error bars represent standard deviation. (B-C) RT-PCR of ELF3, FOXA1, GATA3, GRHL3, IRF1, KLF5 and PPARG mRNA expression by (B) NHB cells and (C) NHU cells. RNA was extracted at the 12, 24 and 48 h time-points and then DNAase-treated and used to generate cDNA for RT-PCR. GAPDH was used as an internal loading control. A no-template (H 2 O) control was included as a negative control for the PCR reaction, and genomic DNA was used as the positive control (+ctrl). No product was amplified from RT-negative controls (not shown). Experiments were performed on n = 2 independent NHB donor cell lines and representative results shown.
Figure Legend Snippet: Comparison of uroplakin and urothelium differentiation-associated transcription factor gene expression by NHB and NHU cell cultures. Employing protocols developed to differentiate NHU cells by PPARγ activation, cell cultures of NHB or NHU cells were exposed to 1 µM troglitazone and 1 µM PD153035 (TZ/PD) for 24 h, maintained in 1 µM PD153035 and harvested at 12, 24, 48 and/or 72 h. Control cultures were exposed to vehicle (0.1% DMSO) alone. (A) RTqPCR for three independent NHB cell lines (represented by different symbols), versus a single NHU cell line for comparison of uroplakin (UPK1A, UPK1B, UPK2, UPK3A and UPK3B) mRNA expression at the 72 h time-point. All data has been normalised to GAPDH expression and is presented relative to the DMSO-treated NHB cells for each gene except UPK3A, where the data is shown relative to the DMSO treated NHU cells due to absent UPK3A gene expression by NHB cells. BLOD = Below Limit of Detection. Statistical analysis was performed using a two-tailed, paired t -test to determine whether TZ/PD resulted in any significant change in gene expression in NHB cells. *represents P ≤ 0.05, ** represents P ≤ 0.01. Error bars represent standard deviation. (B-C) RT-PCR of ELF3, FOXA1, GATA3, GRHL3, IRF1, KLF5 and PPARG mRNA expression by (B) NHB cells and (C) NHU cells. RNA was extracted at the 12, 24 and 48 h time-points and then DNAase-treated and used to generate cDNA for RT-PCR. GAPDH was used as an internal loading control. A no-template (H 2 O) control was included as a negative control for the PCR reaction, and genomic DNA was used as the positive control (+ctrl). No product was amplified from RT-negative controls (not shown). Experiments were performed on n = 2 independent NHB donor cell lines and representative results shown.

Techniques Used: Expressing, Activation Assay, Two Tailed Test, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Negative Control, Polymerase Chain Reaction, Positive Control, Amplification

Evaluation of ELF3 , FOXA1 , GATA3 and PPARG expression in NHB cells. RNA and protein were extracted from parallel cultures of NHB and NHU cells at 72 h following exposure to the PPARγ-activating TZ/PD protocol, or a vehicle control (0.1% DMSO). (A) RTqPCR results combined from three independent NHB cell lines (represented by different symbols), with a single NHU cell line for comparison. All data is normalised to GAPDH expression and is presented relative to the DMSO-treated NHB cell control for each gene. Statistical analysis was performed using a two-tailed, paired t -test to test if TZ/PD treatment resulted in any significant change in gene expression in NHB cells. ** represents P ≤ 0.01. Error bars represent standard deviation. (B) Immunoblot of whole cell protein lysates from representative NHB and NHU cell cultures following exposure to the TZ/PD protocol, 1 µM PD153035 alone, or vehicle (0.1% DMSO) for 72 h. ACTB was included as an internal loading control. Experiments performed on n = 3 independent NHB cell lines with similar results. (C) Immunofluorescence microscopy of ELF3, FOXA1/2, GATA3 and PPARγ in representative NHB and NHU cell cultures. Images taken at identical exposures to demonstrate differences in labelling intensity between the two cell types. Experiments performed on n = 3 independent NHB donor cell lines with similar results. Scale bar ≡ 50 µm.
Figure Legend Snippet: Evaluation of ELF3 , FOXA1 , GATA3 and PPARG expression in NHB cells. RNA and protein were extracted from parallel cultures of NHB and NHU cells at 72 h following exposure to the PPARγ-activating TZ/PD protocol, or a vehicle control (0.1% DMSO). (A) RTqPCR results combined from three independent NHB cell lines (represented by different symbols), with a single NHU cell line for comparison. All data is normalised to GAPDH expression and is presented relative to the DMSO-treated NHB cell control for each gene. Statistical analysis was performed using a two-tailed, paired t -test to test if TZ/PD treatment resulted in any significant change in gene expression in NHB cells. ** represents P ≤ 0.01. Error bars represent standard deviation. (B) Immunoblot of whole cell protein lysates from representative NHB and NHU cell cultures following exposure to the TZ/PD protocol, 1 µM PD153035 alone, or vehicle (0.1% DMSO) for 72 h. ACTB was included as an internal loading control. Experiments performed on n = 3 independent NHB cell lines with similar results. (C) Immunofluorescence microscopy of ELF3, FOXA1/2, GATA3 and PPARγ in representative NHB and NHU cell cultures. Images taken at identical exposures to demonstrate differences in labelling intensity between the two cell types. Experiments performed on n = 3 independent NHB donor cell lines with similar results. Scale bar ≡ 50 µm.

Techniques Used: Expressing, Two Tailed Test, Standard Deviation, Immunofluorescence, Microscopy

37) Product Images from "Evasion of immunosurveillance by genomic alterations of PPARγ/RXRα in bladder cancer"

Article Title: Evasion of immunosurveillance by genomic alterations of PPARγ/RXRα in bladder cancer

Journal: Nature Communications

doi: 10.1038/s41467-017-00147-w

S427F mutation in RXRα stabilizes heterodimerization with PPARγ and promotes the agonistic conformation. a Sizing profile of RXRα S427F mutant ( green ), PPARγ ( purple ), and the heterodimer ( magenta ). Both RXRα S427F and PPARγ run as monomers. When mixed together in 1:1 stoichiometry, the elution profile shifts demonstrating formation of the heterodimer in the absence of ligand. b SPR demonstrating enhanced interaction between RXRα S427F mutant and PPARγ. RXRα was immobilized to the CM5 chip by amine coupling and PPARγ was injected in dose response from 3 μM to 24 nM with 60 s association phase and 120 s disassociation. c Overall crystal structure of the heterodimer complex of RXRα S427F mutant ( green ) and PPARγ ( blue ) with the co-activator peptide Src1 ( red ). The agonists 9-cis-retinoic acid and rosiglitazone are rendered as spheres. The AF-2 helix (Helix H12) of PPARγ has been highlighted in magenta . RXRα S427 and PPARγ Y477 are rendered as sticks and located in the dimer interface. d Zoom in of the heterodimer interface shows the S427F mutation of RXRα ( green ) introduces a π-stacking interaction with Y477 of PPARg ( blue ) at the C-terminus ( magenta ). The 2Fo–Fc electron density map is shown in gray and contoured at 1.2 s
Figure Legend Snippet: S427F mutation in RXRα stabilizes heterodimerization with PPARγ and promotes the agonistic conformation. a Sizing profile of RXRα S427F mutant ( green ), PPARγ ( purple ), and the heterodimer ( magenta ). Both RXRα S427F and PPARγ run as monomers. When mixed together in 1:1 stoichiometry, the elution profile shifts demonstrating formation of the heterodimer in the absence of ligand. b SPR demonstrating enhanced interaction between RXRα S427F mutant and PPARγ. RXRα was immobilized to the CM5 chip by amine coupling and PPARγ was injected in dose response from 3 μM to 24 nM with 60 s association phase and 120 s disassociation. c Overall crystal structure of the heterodimer complex of RXRα S427F mutant ( green ) and PPARγ ( blue ) with the co-activator peptide Src1 ( red ). The agonists 9-cis-retinoic acid and rosiglitazone are rendered as spheres. The AF-2 helix (Helix H12) of PPARγ has been highlighted in magenta . RXRα S427 and PPARγ Y477 are rendered as sticks and located in the dimer interface. d Zoom in of the heterodimer interface shows the S427F mutation of RXRα ( green ) introduces a π-stacking interaction with Y477 of PPARg ( blue ) at the C-terminus ( magenta ). The 2Fo–Fc electron density map is shown in gray and contoured at 1.2 s

Techniques Used: Mutagenesis, SPR Assay, Chromatin Immunoprecipitation, Injection

Tumor-intrinsic activation of PPARγ/RXRα is negatively correlated with immune infiltration. a Pathway enrichment analysis of genes differentially expressed in RXRA-S427Y, RXRA-S427F and PPARG overexpressing T24 lines relative to respective controls. Top suppressed pathways are shown. The analysis was based on three biological replicates. b Dot plot showing expression correlation of all genes with the curated immune signature (refer to “Methods”) vs. correlation with PPARG in bladder tumors ( n = 385) from TCGA. c Heatmap presenting associations between RXRA mutations and PPARG expression with T-cell markers ( top, green label ), immune checkpoint molecules ( middle, yellow label ), and pro-inflammatory factors ( bottom, lavender label ) in TCGA MIBC ( n = 385). d IHC staining of PPARγ and CD8 in two representative human bladder tumor samples from a clinical cohort ( n = 23, Eisai cohort). Scale bars: 100 μm. e Summary of the IHC results of Eisai cohort shown in d . Distribution of CD8+ T-cell infiltration in bladder tumors expressing high (scores 2–4) or low (score 1) levels of PPARγ protein. f Whisker plot representing IHC staining of infiltrating CD8+ T cells and PPARγ protein expression of MIBC samples from the bladder cancer meta-dataset ( n = 118). No expression, score = 1; High expression, score = 4. The bold lines: median; the boxes: interquartile range ( IQR ); the upper whiskers: min(max(x), Q_3 + 1.5 * IQR); the lower whiskers: max(min(x), Q_1−1.5 * IQR). Statistical analysis was performed using Kruskal–Wallis test
Figure Legend Snippet: Tumor-intrinsic activation of PPARγ/RXRα is negatively correlated with immune infiltration. a Pathway enrichment analysis of genes differentially expressed in RXRA-S427Y, RXRA-S427F and PPARG overexpressing T24 lines relative to respective controls. Top suppressed pathways are shown. The analysis was based on three biological replicates. b Dot plot showing expression correlation of all genes with the curated immune signature (refer to “Methods”) vs. correlation with PPARG in bladder tumors ( n = 385) from TCGA. c Heatmap presenting associations between RXRA mutations and PPARG expression with T-cell markers ( top, green label ), immune checkpoint molecules ( middle, yellow label ), and pro-inflammatory factors ( bottom, lavender label ) in TCGA MIBC ( n = 385). d IHC staining of PPARγ and CD8 in two representative human bladder tumor samples from a clinical cohort ( n = 23, Eisai cohort). Scale bars: 100 μm. e Summary of the IHC results of Eisai cohort shown in d . Distribution of CD8+ T-cell infiltration in bladder tumors expressing high (scores 2–4) or low (score 1) levels of PPARγ protein. f Whisker plot representing IHC staining of infiltrating CD8+ T cells and PPARγ protein expression of MIBC samples from the bladder cancer meta-dataset ( n = 118). No expression, score = 1; High expression, score = 4. The bold lines: median; the boxes: interquartile range ( IQR ); the upper whiskers: min(max(x), Q_3 + 1.5 * IQR); the lower whiskers: max(min(x), Q_1−1.5 * IQR). Statistical analysis was performed using Kruskal–Wallis test

Techniques Used: Activation Assay, Expressing, Immunohistochemistry, Staining, Whisker Assay

RXRα S427F/Y functionally promotes ligand-independent PPARγ signaling in human bladder cancer lines. a Heat map representing pathways activated/suppressed in RXRα S427Y , RXRα S427F and PPARγ overexpressing lines relative to their respective controls. Orange represents pathway activation and blue represents pathway suppression. The analysis was based on three biological replicates. b Upper , western blot of RXRα confirming overexpression of RXRα WT (WT), RXRα S427F (S427F) and RXRα S427Y (S427Y) in T24 cells relative to control (Vec). Lower , RT-qPCR analysis of ANGPTL4 and PLIN2 in various engineered lines. c Upper , western blot confirming overexpression of PPARγ in T24 line relative to control (Vec). Lower , RT-qPCR analysis of ANGPTL4 , PLIN2 , ACOX1 and PDK4 in engineered lines. d Upper , western blot of RXRα and PPARγ in SV-HUC line engineered to inducibly overexpress RXRα S427F and knockdown PPARγ by multiple shRNAs (sh#4, 5 and 9) upon doxycycline ( DOX ) treatment. Lower , RT-qPCR analysis of PLIN2 , ACOX1 and PSCA in various SV-HUC-1 engineered lines. +/− represents presence or absence of DOX treatment respectively. e RT-qPCR analysis of ANGPTL4 and PLIN2 in HT-1197 (carrying endogenous RXRA S427F ), 5637 and UM-UC9 (PPARG amplified) lines treated with DMSO or T0070907 for 24 h. All RT-qPCR data is normalized to GAPDH and presented as mean fold change vs. control ± SEM of at least three biological replicates
Figure Legend Snippet: RXRα S427F/Y functionally promotes ligand-independent PPARγ signaling in human bladder cancer lines. a Heat map representing pathways activated/suppressed in RXRα S427Y , RXRα S427F and PPARγ overexpressing lines relative to their respective controls. Orange represents pathway activation and blue represents pathway suppression. The analysis was based on three biological replicates. b Upper , western blot of RXRα confirming overexpression of RXRα WT (WT), RXRα S427F (S427F) and RXRα S427Y (S427Y) in T24 cells relative to control (Vec). Lower , RT-qPCR analysis of ANGPTL4 and PLIN2 in various engineered lines. c Upper , western blot confirming overexpression of PPARγ in T24 line relative to control (Vec). Lower , RT-qPCR analysis of ANGPTL4 , PLIN2 , ACOX1 and PDK4 in engineered lines. d Upper , western blot of RXRα and PPARγ in SV-HUC line engineered to inducibly overexpress RXRα S427F and knockdown PPARγ by multiple shRNAs (sh#4, 5 and 9) upon doxycycline ( DOX ) treatment. Lower , RT-qPCR analysis of PLIN2 , ACOX1 and PSCA in various SV-HUC-1 engineered lines. +/− represents presence or absence of DOX treatment respectively. e RT-qPCR analysis of ANGPTL4 and PLIN2 in HT-1197 (carrying endogenous RXRA S427F ), 5637 and UM-UC9 (PPARG amplified) lines treated with DMSO or T0070907 for 24 h. All RT-qPCR data is normalized to GAPDH and presented as mean fold change vs. control ± SEM of at least three biological replicates

Techniques Used: Activation Assay, Western Blot, Over Expression, Quantitative RT-PCR, Amplification

PPARγ/RXRα S427F confers partial resistance to immunotherapies. a RT-qPCR analysis of chemokines/cytokines in T24 lines engineered to overexpress RXRA-WT, RXRA-S427F, RXRA-S427Y ( upper ), and PPARG ( lower ). Controls are RXRA-WT for RXRA mutant lines and vector control (Vec) for PPARG overexpressing line. Expression normalized to GAPDH and data presented as mean fold change vs. control ± SEM of three biological replicates. b Chemokine array analysis of conditioned media collected from T24 lines engineered to overexpress PPARG (PPARγ) vs. control (Vec). Dotted boxes represent controls. Significant changes in secretion are outlined. One representative of three independent experiments is shown. c FACS based quantitation of infiltrating CD3 + CD8 + double positive T cells into subcutaneously implanted MBT2 tumors overexpressing RXRA-WT ( n = 6) or RXRA-S427F ( n = 6). Data presented as percent of total tumor-derived cells following dissociation. d Left , individual MBT2-RXRα WT tumor volumes in response to PBS ( red , n = 12) or anti-CTLA4 ( blue , n = 12). P = 0.0189 at day 7. Right , individual MBT2-RXRα S427F tumor volumes in response to PBS ( red , n = 12) or anti-CTLA4 ( blue , n = 12). P > 0.05 at day 7. One-way ANOVA followed by Tukey’s post-hoc test performed. e Heatmap presenting pathway level analysis (activation, red ; suppression, blue ) of differentially expressed genes in PPARγ knockdown lines (PPARγ-sh#4 and -sh#9 engineered in SV-HUC-1 line expressing RXRA-S427Y) relative to vector control. The analysis was based on three biological replicates. f Left , Knockdown of PPARγ or RXRα by shRNAs in HT-1197 cells. GAPDH was used as the control. Right , RT-qPCR analysis of inflammatory genes CCL2 and CXCL10 following inducible knockdown of PPARγ and RXRα in HT-1197 cells. Data normalized to GAPDH and presented as mean fold change (Dox treated vs. untreated) ± SEM of three biological replicates. g RT-qPCR analysis of IL8 and CCL2 following treatment with PPARγ agonist rosiglitazone (Rosi) or PPARγ antagonist T0070907 in 5637 cells. Data normalized to GAPDH and presented as mean fold change ± SEM of three biological replicates. h Schematic representation of the role of tumor-intrinsic PPARγ/RXRα S427F/Y in transcriptional regulation and immunosurveillance. CoA, co-activator complex; CoR, co-repressor complex; ITF, inflammation-related transcription factors
Figure Legend Snippet: PPARγ/RXRα S427F confers partial resistance to immunotherapies. a RT-qPCR analysis of chemokines/cytokines in T24 lines engineered to overexpress RXRA-WT, RXRA-S427F, RXRA-S427Y ( upper ), and PPARG ( lower ). Controls are RXRA-WT for RXRA mutant lines and vector control (Vec) for PPARG overexpressing line. Expression normalized to GAPDH and data presented as mean fold change vs. control ± SEM of three biological replicates. b Chemokine array analysis of conditioned media collected from T24 lines engineered to overexpress PPARG (PPARγ) vs. control (Vec). Dotted boxes represent controls. Significant changes in secretion are outlined. One representative of three independent experiments is shown. c FACS based quantitation of infiltrating CD3 + CD8 + double positive T cells into subcutaneously implanted MBT2 tumors overexpressing RXRA-WT ( n = 6) or RXRA-S427F ( n = 6). Data presented as percent of total tumor-derived cells following dissociation. d Left , individual MBT2-RXRα WT tumor volumes in response to PBS ( red , n = 12) or anti-CTLA4 ( blue , n = 12). P = 0.0189 at day 7. Right , individual MBT2-RXRα S427F tumor volumes in response to PBS ( red , n = 12) or anti-CTLA4 ( blue , n = 12). P > 0.05 at day 7. One-way ANOVA followed by Tukey’s post-hoc test performed. e Heatmap presenting pathway level analysis (activation, red ; suppression, blue ) of differentially expressed genes in PPARγ knockdown lines (PPARγ-sh#4 and -sh#9 engineered in SV-HUC-1 line expressing RXRA-S427Y) relative to vector control. The analysis was based on three biological replicates. f Left , Knockdown of PPARγ or RXRα by shRNAs in HT-1197 cells. GAPDH was used as the control. Right , RT-qPCR analysis of inflammatory genes CCL2 and CXCL10 following inducible knockdown of PPARγ and RXRα in HT-1197 cells. Data normalized to GAPDH and presented as mean fold change (Dox treated vs. untreated) ± SEM of three biological replicates. g RT-qPCR analysis of IL8 and CCL2 following treatment with PPARγ agonist rosiglitazone (Rosi) or PPARγ antagonist T0070907 in 5637 cells. Data normalized to GAPDH and presented as mean fold change ± SEM of three biological replicates. h Schematic representation of the role of tumor-intrinsic PPARγ/RXRα S427F/Y in transcriptional regulation and immunosurveillance. CoA, co-activator complex; CoR, co-repressor complex; ITF, inflammation-related transcription factors

Techniques Used: Quantitative RT-PCR, Mutagenesis, Plasmid Preparation, Expressing, FACS, Quantitation Assay, Derivative Assay, Activation Assay

38) Product Images from "Ilex latifolia Thunb protects mice from HFD-induced body weight gain"

Article Title: Ilex latifolia Thunb protects mice from HFD-induced body weight gain

Journal: Scientific Reports

doi: 10.1038/s41598-017-15292-x

The expression of lipogenic genes in liver and adipose tissues. ( a ) mRNA expression levels of Pparg(Ppar γ ) , Fas , C/ebpα and Srebp1c in liver. ( b ) mRNA expression levels of Pparg , Fas , C/ebpα and Srebp1c in eWAT. ( c ) PPARγ, p-SREBP1c, SREBP-1c, p-AMPK and AMPK levels in liver. ( d ) PPARγ, p-SREBP1c, SREBP-1c, p-AMPK and AMPK levels in eWAT. Full-length blots are presented in Supplementary Figure S7 . n = 5 for each group. Significant differences between ND and HFD are indicated as ### P
Figure Legend Snippet: The expression of lipogenic genes in liver and adipose tissues. ( a ) mRNA expression levels of Pparg(Ppar γ ) , Fas , C/ebpα and Srebp1c in liver. ( b ) mRNA expression levels of Pparg , Fas , C/ebpα and Srebp1c in eWAT. ( c ) PPARγ, p-SREBP1c, SREBP-1c, p-AMPK and AMPK levels in liver. ( d ) PPARγ, p-SREBP1c, SREBP-1c, p-AMPK and AMPK levels in eWAT. Full-length blots are presented in Supplementary Figure S7 . n = 5 for each group. Significant differences between ND and HFD are indicated as ### P

Techniques Used: Expressing

39) Product Images from "Adipogenesis and insulin sensitivity in obesity are regulated by retinoid-related orphan receptor gamma"

Article Title: Adipogenesis and insulin sensitivity in obesity are regulated by retinoid-related orphan receptor gamma

Journal: EMBO Molecular Medicine

doi: 10.1002/emmm.201100172

RORγ inhibits expression and activity of differentiation genes A. Western blot of nuclear extracts (RORγ, PPARγ, C/EBPα, C/EBPβ and C/EBPδ) or cytosolic extracts (MMP3, PREF-1, LPL and Adiponectin) during differentiation of 3T3-L1 cells overexpressing RORγ or empty vector. Differentiation without rosiglitazone was induced at time point 0 h. Lamin B and γ-Tubulin were used as loading controls. B,C,D. qPCR for the adipogenic genes c-Jun, c-Fos and A-Fabp during differentiation of 3T3-L1 cells overexpressing RORγ or empty vector ( n = 4). E. Firefly luciferase activity of the PPARγ response element (PPRE) normalized to renilla luciferase before, 1 and 2 days after induction of differentiation in 3T3-L1 cells overexpressing RORγ or empty vector ( n = 4). F. Firefly luciferase activity of PPARγ promoter normalized to renilla luciferase in 3T3-L1 cells overexpressing RORγ or empty vector during differentiation at indicated time points ( n = 8). The results shown are representative of three independent experiments. Values represent means ± SD, * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001.
Figure Legend Snippet: RORγ inhibits expression and activity of differentiation genes A. Western blot of nuclear extracts (RORγ, PPARγ, C/EBPα, C/EBPβ and C/EBPδ) or cytosolic extracts (MMP3, PREF-1, LPL and Adiponectin) during differentiation of 3T3-L1 cells overexpressing RORγ or empty vector. Differentiation without rosiglitazone was induced at time point 0 h. Lamin B and γ-Tubulin were used as loading controls. B,C,D. qPCR for the adipogenic genes c-Jun, c-Fos and A-Fabp during differentiation of 3T3-L1 cells overexpressing RORγ or empty vector ( n = 4). E. Firefly luciferase activity of the PPARγ response element (PPRE) normalized to renilla luciferase before, 1 and 2 days after induction of differentiation in 3T3-L1 cells overexpressing RORγ or empty vector ( n = 4). F. Firefly luciferase activity of PPARγ promoter normalized to renilla luciferase in 3T3-L1 cells overexpressing RORγ or empty vector during differentiation at indicated time points ( n = 8). The results shown are representative of three independent experiments. Values represent means ± SD, * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001.

Techniques Used: Expressing, Activity Assay, Western Blot, Plasmid Preparation, Real-time Polymerase Chain Reaction, Luciferase

40) Product Images from "Inhibition of fatty acid synthase is protective in pulmonary hypertension) Inhibition of fatty acid synthase is protective in pulmonary hypertension"

Article Title: Inhibition of fatty acid synthase is protective in pulmonary hypertension) Inhibition of fatty acid synthase is protective in pulmonary hypertension

Journal: British Journal of Pharmacology

doi: 10.1111/bph.13495

The mitochondrial dysfunction and insulin resistance in hypoxic HPASMCs. (A) Representative fluorescence‐activated cell sorter (FACS) dot plots of HPASMCs stained with mitosox. The red quadrants represent HPASMCs positive for mitosox ( n = 6 in each group); (B) representative picture of TMRM staining in HPASMCs exposed to hypoxia and treated with either FAS siRNA or scrambled siRNA (5×, n = 5 in each group); and (C) treatment of FAS siRNA improves the insulin resistance. Quantification of pAMPK/AMPK, pAkt/Akt and PPARγ/actin ratios and a representative western blot in primary cultures of HPASMCs ( n = 6 in each group, except hypoxia + Scr.siRNA group only where n = 3). + P
Figure Legend Snippet: The mitochondrial dysfunction and insulin resistance in hypoxic HPASMCs. (A) Representative fluorescence‐activated cell sorter (FACS) dot plots of HPASMCs stained with mitosox. The red quadrants represent HPASMCs positive for mitosox ( n = 6 in each group); (B) representative picture of TMRM staining in HPASMCs exposed to hypoxia and treated with either FAS siRNA or scrambled siRNA (5×, n = 5 in each group); and (C) treatment of FAS siRNA improves the insulin resistance. Quantification of pAMPK/AMPK, pAkt/Akt and PPARγ/actin ratios and a representative western blot in primary cultures of HPASMCs ( n = 6 in each group, except hypoxia + Scr.siRNA group only where n = 3). + P

Techniques Used: Fluorescence, FACS, Staining, Western Blot

41) Product Images from "Magnesium ion influx reduces neuroinflammation in Aβ precursor protein/Presenilin 1 transgenic mice by suppressing the expression of interleukin-1β"

Article Title: Magnesium ion influx reduces neuroinflammation in Aβ precursor protein/Presenilin 1 transgenic mice by suppressing the expression of interleukin-1β

Journal: Cellular and Molecular Immunology

doi: 10.1038/cmi.2015.93

Proposed cascade of the signaling events regulating the expression of IL-1β by MgT. In detail, attenuated levels of Mg 2+ in APP/PS1 transgenic mice will elevate the production of IL-1β and Aβ via ERK1/2- and PPARγ-dependent pathways in glial cells of the APP/PS1 transgenic mouse brain, which in turn will potentially regulate the pathogenesis of AD. Interestingly, the highly secreted IL-1β and Aβ in the CSF are able to regulate the synthesis of IL-1β in the cerebral cortex of APP/PS1 mice. These observations might be instrumental for understanding the roles of Mg 2+ in suppressing the neuroinflammation of AD.
Figure Legend Snippet: Proposed cascade of the signaling events regulating the expression of IL-1β by MgT. In detail, attenuated levels of Mg 2+ in APP/PS1 transgenic mice will elevate the production of IL-1β and Aβ via ERK1/2- and PPARγ-dependent pathways in glial cells of the APP/PS1 transgenic mouse brain, which in turn will potentially regulate the pathogenesis of AD. Interestingly, the highly secreted IL-1β and Aβ in the CSF are able to regulate the synthesis of IL-1β in the cerebral cortex of APP/PS1 mice. These observations might be instrumental for understanding the roles of Mg 2+ in suppressing the neuroinflammation of AD.

Techniques Used: Expressing, Transgenic Assay, Mouse Assay

Involvement of ERK1/2 and PPARγ pathways in regulating the expression of IL-1β in MgT-treated A172 or D1A cells. Human glioblastoma A172 ( a ) or mouse astrocytes/microglia D1A cells ( b ) were treated with MgT (50 μM) for 48 h. In select experiments, A172 cells were treated with PD98059 (10 μM) in the absence or presence of MgT (50 μM) for 48 h ( c and e ). In separate experiments, the cells were transfected with ERK1/2 ( d ) or PPARγ siRNA ( f ) before incubation with MgT (50 μM) for 48 h. In distinct experiments, A172 cells were treated with GW9662 (1 μM) in the absence or presence of MgT (50 μM) for 48 h ( e '). In other experiments, primary cultured astrocytes were treated with MgT (50 μM) in the absence or presence of PD98059 (10 μM) (g) or GW9662 (1 μM) for 48 h (h). Total ERK1/2 ( c , d and g upper panel), phosphorylated ERK1/2 levels ( c and g upper panel), total PPARγ ( e and f upper panel), and phosphorylated PPARγ ( e ) were detected by immunoblotting using specific Abs. Equal lane loading is demonstrated by the similar intensities of total β-actin. IL-1β protein and mRNA levels were determined by IL-1β enzyme immunoassay kits and qRT-PCR, respectively. The total amounts of protein and GAPDH served as internal controls. The data represent the means ± SE of three independent experiments. * P
Figure Legend Snippet: Involvement of ERK1/2 and PPARγ pathways in regulating the expression of IL-1β in MgT-treated A172 or D1A cells. Human glioblastoma A172 ( a ) or mouse astrocytes/microglia D1A cells ( b ) were treated with MgT (50 μM) for 48 h. In select experiments, A172 cells were treated with PD98059 (10 μM) in the absence or presence of MgT (50 μM) for 48 h ( c and e ). In separate experiments, the cells were transfected with ERK1/2 ( d ) or PPARγ siRNA ( f ) before incubation with MgT (50 μM) for 48 h. In distinct experiments, A172 cells were treated with GW9662 (1 μM) in the absence or presence of MgT (50 μM) for 48 h ( e '). In other experiments, primary cultured astrocytes were treated with MgT (50 μM) in the absence or presence of PD98059 (10 μM) (g) or GW9662 (1 μM) for 48 h (h). Total ERK1/2 ( c , d and g upper panel), phosphorylated ERK1/2 levels ( c and g upper panel), total PPARγ ( e and f upper panel), and phosphorylated PPARγ ( e ) were detected by immunoblotting using specific Abs. Equal lane loading is demonstrated by the similar intensities of total β-actin. IL-1β protein and mRNA levels were determined by IL-1β enzyme immunoassay kits and qRT-PCR, respectively. The total amounts of protein and GAPDH served as internal controls. The data represent the means ± SE of three independent experiments. * P

Techniques Used: Expressing, Transfection, Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

42) Product Images from "Obesity Reduces Bone Density Associated with Activation of PPAR? and Suppression of Wnt/?-Catenin in Rapidly Growing Male Rats"

Article Title: Obesity Reduces Bone Density Associated with Activation of PPAR? and Suppression of Wnt/?-Catenin in Rapidly Growing Male Rats

Journal: PLoS ONE

doi: 10.1371/journal.pone.0013704

Serum from HFD-induced obese rats and an artificial FA mixture suppress osteoblast differentiation. (A), ST2 cells were cultured in 12 well plates. Cells were treated with 2% serum from LFD, MFD or HFD rats, 50 ng/ml Wnt3a, 400 µM FAs and their combination for 7 days in the presence or absence of osteogenic medium. Alkaline phophatase staining was performed. (B), 2% serum from HFD-induced obese rats and an artificial FA mixture significantly decreased TCF/LEF-dependent transcription of a luciferase reporter gene (TOPFLASH) in C2C12 osteoblastic cells compared with cells treated with LFD serum. Luciferase activity in C2C12 cells transfected with a PPRE-luc reporter construct and treated with 2% serum from LFD, MFD or HFD-fed rats, 50 ng/ml Wnt3a, 400 µM FAs and their combination for 24 h. (C), β-catenin gene was knock down using β-catenin siRNA in ST2 cells. After 24 h of β-catenin siRNA, cell proteins were collected and western blot was performed for β-catenin and PPARγ. Bars are expressed as mean ± SEM in triplicates. *, P
Figure Legend Snippet: Serum from HFD-induced obese rats and an artificial FA mixture suppress osteoblast differentiation. (A), ST2 cells were cultured in 12 well plates. Cells were treated with 2% serum from LFD, MFD or HFD rats, 50 ng/ml Wnt3a, 400 µM FAs and their combination for 7 days in the presence or absence of osteogenic medium. Alkaline phophatase staining was performed. (B), 2% serum from HFD-induced obese rats and an artificial FA mixture significantly decreased TCF/LEF-dependent transcription of a luciferase reporter gene (TOPFLASH) in C2C12 osteoblastic cells compared with cells treated with LFD serum. Luciferase activity in C2C12 cells transfected with a PPRE-luc reporter construct and treated with 2% serum from LFD, MFD or HFD-fed rats, 50 ng/ml Wnt3a, 400 µM FAs and their combination for 24 h. (C), β-catenin gene was knock down using β-catenin siRNA in ST2 cells. After 24 h of β-catenin siRNA, cell proteins were collected and western blot was performed for β-catenin and PPARγ. Bars are expressed as mean ± SEM in triplicates. *, P

Techniques Used: Cell Culture, Staining, Luciferase, Activity Assay, Transfection, Construct, Western Blot

An artificial NEFA mixture based on the ratio and concentrations of FAs appearing in serum from obese rats reciprocally regulates β-catenin and PPARγ expression in ST2 cells. (A), GC-MS analysis of NEFA. To measure NEFA composition, 100 µl of serum from LFD, MFD or HFD rats was characterized and quantified by a Shimazu QP-2010 GC-MS system after TLC separation. (B), Mean concentrations of the 5 major FAs in NEFA are presented. (C), ST2 cells were treated with a NEFA mixture based on individual FA concentrations appearing in serum in LFD, MFD and HFD rats for 24 and 48 h. Real-time RT-PCR of β-catenin and PPARγ mRNA expressions. (D) Western blot analysis of β-catenin and PPARγ (n = 3). LFD, low fat diet (control pelleted AIN-93G 14% fat diet); MFD, medium fat TEN diet (25% fat diet); HFD, high fat TEN diet (45% fat diet). Data bars are expressed as mean ± SEM (n = 3/treatment). *, P
Figure Legend Snippet: An artificial NEFA mixture based on the ratio and concentrations of FAs appearing in serum from obese rats reciprocally regulates β-catenin and PPARγ expression in ST2 cells. (A), GC-MS analysis of NEFA. To measure NEFA composition, 100 µl of serum from LFD, MFD or HFD rats was characterized and quantified by a Shimazu QP-2010 GC-MS system after TLC separation. (B), Mean concentrations of the 5 major FAs in NEFA are presented. (C), ST2 cells were treated with a NEFA mixture based on individual FA concentrations appearing in serum in LFD, MFD and HFD rats for 24 and 48 h. Real-time RT-PCR of β-catenin and PPARγ mRNA expressions. (D) Western blot analysis of β-catenin and PPARγ (n = 3). LFD, low fat diet (control pelleted AIN-93G 14% fat diet); MFD, medium fat TEN diet (25% fat diet); HFD, high fat TEN diet (45% fat diet). Data bars are expressed as mean ± SEM (n = 3/treatment). *, P

Techniques Used: Expressing, Gas Chromatography-Mass Spectrometry, Thin Layer Chromatography, Quantitative RT-PCR, Western Blot

Serum from HFD fed rats and an artificial FA mixture down-regulate β-catenin but up-regulate PPARγ in ST2 cells, Wnt3a does the opposite. (A), ST2 cells were treated with 2% serum from LFD, MFD or HFD rats for 3 days. Cell RNAs were isolated and real-time PCR was performed for β-catenin and PPARγ. (B), ST2 cells were treated with three different concentrations of FAs (200, 400 or 800 µM) for 24 h and 48 h. Cell RNAs were isolated and real-time PCR was performed for β-batenin and PPARγ. (C), ST2 cells were treated with a FA mixture and Wnt3a respectively for 24 h. Cell protein lysates were collected, and western blots were performed for β-catenin, PPARγ and β-actin in duplicates. Data bars are expressed as mean ± SEM (n = 3/treatment). Means with different letters differ significantly from each other at P
Figure Legend Snippet: Serum from HFD fed rats and an artificial FA mixture down-regulate β-catenin but up-regulate PPARγ in ST2 cells, Wnt3a does the opposite. (A), ST2 cells were treated with 2% serum from LFD, MFD or HFD rats for 3 days. Cell RNAs were isolated and real-time PCR was performed for β-catenin and PPARγ. (B), ST2 cells were treated with three different concentrations of FAs (200, 400 or 800 µM) for 24 h and 48 h. Cell RNAs were isolated and real-time PCR was performed for β-batenin and PPARγ. (C), ST2 cells were treated with a FA mixture and Wnt3a respectively for 24 h. Cell protein lysates were collected, and western blots were performed for β-catenin, PPARγ and β-actin in duplicates. Data bars are expressed as mean ± SEM (n = 3/treatment). Means with different letters differ significantly from each other at P

Techniques Used: Isolation, Real-time Polymerase Chain Reaction, Western Blot

Activation of PPARγ and regulation of its target gene. (A), nuclear proteins were extracted from rat tibia and from ST2 cells treated with rat serum and FAs mixture using 10 cm dish. TransAM was performed for activity of PPARγ. (B) ST2 cells were treated for 24 h with FAs mixture according to the concentrations that appeared in animal circulation. ChIP of mouse aP2 enhancer elements by specific anti PPARγ antibody. Control, normal cell culture medium; LFD, low fat diet (control pelleted AIN-93G 14% fat diet); MFD, medium fat TEN diet (25% fat diet); HFD, high fat TEN diet (45% fat diet). Bars are expressed as mean ± SEM in triplicates. *, P
Figure Legend Snippet: Activation of PPARγ and regulation of its target gene. (A), nuclear proteins were extracted from rat tibia and from ST2 cells treated with rat serum and FAs mixture using 10 cm dish. TransAM was performed for activity of PPARγ. (B) ST2 cells were treated for 24 h with FAs mixture according to the concentrations that appeared in animal circulation. ChIP of mouse aP2 enhancer elements by specific anti PPARγ antibody. Control, normal cell culture medium; LFD, low fat diet (control pelleted AIN-93G 14% fat diet); MFD, medium fat TEN diet (25% fat diet); HFD, high fat TEN diet (45% fat diet). Bars are expressed as mean ± SEM in triplicates. *, P

Techniques Used: Activation Assay, Activity Assay, Chromatin Immunoprecipitation, Cell Culture

Increased adipogenesis in bone and bone marrow in HFD-induced obese animals. (A), Represented H E staining picture of increased bone marrow adiposity in tibial bone section from HFD-induced obese animals (10x). (B) and (C), osteocalcin and Runx2 gene expression measured using real-time RT-PCR. (D) and (E), PPARγ and aP2 gene expression measured using real time RT-PCR. Total RNA was isolated from femur of each animal after bone marrow aspiration for RT-PCR. LFD, low fat diet (control pelleted AIN-93G 14% fat diet); MFD, medium fat diet (25% fat diet); HFD, high fat TEN diet (45% fat diet). Data bars are expressed as mean ± SEM (n = 6/group). Means with different letters differ significantly from each other at p
Figure Legend Snippet: Increased adipogenesis in bone and bone marrow in HFD-induced obese animals. (A), Represented H E staining picture of increased bone marrow adiposity in tibial bone section from HFD-induced obese animals (10x). (B) and (C), osteocalcin and Runx2 gene expression measured using real-time RT-PCR. (D) and (E), PPARγ and aP2 gene expression measured using real time RT-PCR. Total RNA was isolated from femur of each animal after bone marrow aspiration for RT-PCR. LFD, low fat diet (control pelleted AIN-93G 14% fat diet); MFD, medium fat diet (25% fat diet); HFD, high fat TEN diet (45% fat diet). Data bars are expressed as mean ± SEM (n = 6/group). Means with different letters differ significantly from each other at p

Techniques Used: Staining, Expressing, Quantitative RT-PCR, Isolation, Reverse Transcription Polymerase Chain Reaction

Increased PPARγ but decreased β-catenin protein expression in bone from HFD-induced obese rats. (A) Representative pictures of immunostained tibial bone sections using an anti-mouse β-catenin polyclonal antibody. White arrows indicate immunostained β-catenin, and black arrows indicate bone tissues. (B), Western blots of PPARγ (from total protein lysates, nuclear and cytoplasmic fractions), β-catenin and β-actin are depicted for four samples from LFD, MFD and HFD groups. LFD, low fat diet (control pelleted AIN-93G 14% fat diet); MFD, medium fat TEN diet (25% fat diet); HFD, high fat TEN diet (45% fat diet). Proteins were isolated from long bone after aspiration of bone marrow cells.
Figure Legend Snippet: Increased PPARγ but decreased β-catenin protein expression in bone from HFD-induced obese rats. (A) Representative pictures of immunostained tibial bone sections using an anti-mouse β-catenin polyclonal antibody. White arrows indicate immunostained β-catenin, and black arrows indicate bone tissues. (B), Western blots of PPARγ (from total protein lysates, nuclear and cytoplasmic fractions), β-catenin and β-actin are depicted for four samples from LFD, MFD and HFD groups. LFD, low fat diet (control pelleted AIN-93G 14% fat diet); MFD, medium fat TEN diet (25% fat diet); HFD, high fat TEN diet (45% fat diet). Proteins were isolated from long bone after aspiration of bone marrow cells.

Techniques Used: Expressing, Western Blot, Isolation

43) Product Images from "The Ras Inhibitors Caveolin-1 and Docking Protein 1 Activate Peroxisome Proliferator-Activated Receptor ? through Spatial Relocalization at Helix 7 of Its Ligand-Binding Domain ▿"

Article Title: The Ras Inhibitors Caveolin-1 and Docking Protein 1 Activate Peroxisome Proliferator-Activated Receptor ? through Spatial Relocalization at Helix 7 of Its Ligand-Binding Domain ▿

Journal: Molecular and Cellular Biology

doi: 10.1128/MCB.01421-10

Endogenous Cav1 promotes nuclear translocation and ligand-dependent transcription of PPARγ target genes. (A) mRNA expression. MKN45 clones were treated for 16 h with vehicle or 1 μM rosiglitazone. C T values from RT-qPCRs were normalized
Figure Legend Snippet: Endogenous Cav1 promotes nuclear translocation and ligand-dependent transcription of PPARγ target genes. (A) mRNA expression. MKN45 clones were treated for 16 h with vehicle or 1 μM rosiglitazone. C T values from RT-qPCRs were normalized

Techniques Used: Translocation Assay, Expressing, Clone Assay

Helix 7 in the LBD of PPARγ harbors a Cav1-binding motif. (A) Mapping of the PPARγ-Cav1 interaction site by CoIP. (Top) Human peptide sequences within the putative docking interfaces of helix 7 (H7) in the PPARγ LBD (aa 350 to
Figure Legend Snippet: Helix 7 in the LBD of PPARγ harbors a Cav1-binding motif. (A) Mapping of the PPARγ-Cav1 interaction site by CoIP. (Top) Human peptide sequences within the putative docking interfaces of helix 7 (H7) in the PPARγ LBD (aa 350 to

Techniques Used: Binding Assay, Co-Immunoprecipitation Assay

Localization of PPARγ in human GC patient tissues and cell lines. (A) IHC on paraffin sections using the polyclonal Ab for PPARγ (brown) and the monoclonal Ab for Cav1 (red). NT, normal stomach; IM, intestinal metaplasia in intestinal-type
Figure Legend Snippet: Localization of PPARγ in human GC patient tissues and cell lines. (A) IHC on paraffin sections using the polyclonal Ab for PPARγ (brown) and the monoclonal Ab for Cav1 (red). NT, normal stomach; IM, intestinal metaplasia in intestinal-type

Techniques Used: Immunohistochemistry

Dok1 augments the ligand-dependent transcriptional activity of PPARγ. (A) Immunofluorescence microscopy of Dok1 showing nuclear translocation of endogenous PPARγ in MKN45 cells but not in AGS cells. Parental GC cells were deprived of serum
Figure Legend Snippet: Dok1 augments the ligand-dependent transcriptional activity of PPARγ. (A) Immunofluorescence microscopy of Dok1 showing nuclear translocation of endogenous PPARγ in MKN45 cells but not in AGS cells. Parental GC cells were deprived of serum

Techniques Used: Activity Assay, Immunofluorescence, Microscopy, Translocation Assay

PPARγ activation inhibits proliferation and upregulates Dok1 in a murine model of GC. (A) Reduced proliferation in pyloric tumor areas of stomachs from CEA424-SV40 T-antigen (Tag) mice upon a 6-week chow diet enriched with 0.02% (wt/wt) rosiglitazone
Figure Legend Snippet: PPARγ activation inhibits proliferation and upregulates Dok1 in a murine model of GC. (A) Reduced proliferation in pyloric tumor areas of stomachs from CEA424-SV40 T-antigen (Tag) mice upon a 6-week chow diet enriched with 0.02% (wt/wt) rosiglitazone

Techniques Used: Activation Assay, Mouse Assay

Model of PPARγ interactions with Ras/MAPK inhibitors in human GC. (A) Cav1 in (K-Ras mutated) AGS cells acts as a sequestor for PPARγ, inhibiting basal and ligand-dependent transcription of PPARγ target genes. Loss of Cav1 leads
Figure Legend Snippet: Model of PPARγ interactions with Ras/MAPK inhibitors in human GC. (A) Cav1 in (K-Ras mutated) AGS cells acts as a sequestor for PPARγ, inhibiting basal and ligand-dependent transcription of PPARγ target genes. Loss of Cav1 leads

Techniques Used:

Loss of Cav1 promotes nuclear localization and ERK-dependent phosphorylation of PPARγ at serine 84. (A, B) Subcellular fractionation. Serum-deprived AGS clones were stimulated for 60 min with TPA (100 nM) or rosiglitazone (10 μM). Representative
Figure Legend Snippet: Loss of Cav1 promotes nuclear localization and ERK-dependent phosphorylation of PPARγ at serine 84. (A, B) Subcellular fractionation. Serum-deprived AGS clones were stimulated for 60 min with TPA (100 nM) or rosiglitazone (10 μM). Representative

Techniques Used: Fractionation, Clone Assay

PPARγ interacts with MEK1 and Cav1 in the cytosol. (A) PPARγ-MEK1 interaction. Serum-deprived AGS clones were stimulated for 30 min with TPA (100 nM) and rosiglitazone (10 μM) before CoIP (IP) from cytosolic lysates using the polyclonal
Figure Legend Snippet: PPARγ interacts with MEK1 and Cav1 in the cytosol. (A) PPARγ-MEK1 interaction. Serum-deprived AGS clones were stimulated for 30 min with TPA (100 nM) and rosiglitazone (10 μM) before CoIP (IP) from cytosolic lysates using the polyclonal

Techniques Used: Clone Assay, Co-Immunoprecipitation Assay

44) Product Images from "Peroxisome Proliferator Activated Receptor gamma (PPARγ) Agonist Rosiglitazone Ameliorate Airway Inflammation by Inhibiting Toll-Like Receptor 2 (TLR2)/Nod-Like Receptor with Pyrin Domain Containing 3 (NLRP3) Inflammatory Corpuscle Activation in Asthmatic Mice"

Article Title: Peroxisome Proliferator Activated Receptor gamma (PPARγ) Agonist Rosiglitazone Ameliorate Airway Inflammation by Inhibiting Toll-Like Receptor 2 (TLR2)/Nod-Like Receptor with Pyrin Domain Containing 3 (NLRP3) Inflammatory Corpuscle Activation in Asthmatic Mice

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

doi: 10.12659/MSM.910766

Expression of toll-like receptor 2 (TLR2) and peroxisome proliferator activated receptor agonist (PPARγ) proteins were significantly decreased after rosiglitazone treatment. ( A ) Expression of TLR2 and PPARγ proteins by western blot analysis. ( B ) Quantitative analysis of protein test strips. Data are represented as the mean ±SD. * P
Figure Legend Snippet: Expression of toll-like receptor 2 (TLR2) and peroxisome proliferator activated receptor agonist (PPARγ) proteins were significantly decreased after rosiglitazone treatment. ( A ) Expression of TLR2 and PPARγ proteins by western blot analysis. ( B ) Quantitative analysis of protein test strips. Data are represented as the mean ±SD. * P

Techniques Used: Expressing, Western Blot

45) Product Images from "Palmitate induces fat accumulation by activating C/EBPβ-mediated G0S2 expression in HepG2 cells"

Article Title: Palmitate induces fat accumulation by activating C/EBPβ-mediated G0S2 expression in HepG2 cells

Journal: World Journal of Gastroenterology

doi: 10.3748/wjg.v23.i43.7705

Palmitate-induced expression of C/EBPβ, PPARγ, and PPARγ target genes in HepG2 cells. A: Palmitate increased mRNA expression of C/EBPβ, PPARγ, and PPARγ target genes ( G0S2 , GPR81 , GPR109A , and Adipoq ) in a dose-dependent manner. mRNA was measured by qPCR. At least three independent experiments were conducted for each measurement. Data are presented as means ± SE. a P
Figure Legend Snippet: Palmitate-induced expression of C/EBPβ, PPARγ, and PPARγ target genes in HepG2 cells. A: Palmitate increased mRNA expression of C/EBPβ, PPARγ, and PPARγ target genes ( G0S2 , GPR81 , GPR109A , and Adipoq ) in a dose-dependent manner. mRNA was measured by qPCR. At least three independent experiments were conducted for each measurement. Data are presented as means ± SE. a P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction

The effects of G0S2 knockdown on C/EBP β, PPAR γ and PPARγ target genes expression, lipid accumulation, and lipolysis in palmitate-treated HepG2 cells. A: HepG2 cells were transfected with control siRNA or G0S2 siRNA, and G0S2 expression was measured by Western blotting. At least three independent experiments were conducted. Data are presented as means ± SE. f P
Figure Legend Snippet: The effects of G0S2 knockdown on C/EBP β, PPAR γ and PPARγ target genes expression, lipid accumulation, and lipolysis in palmitate-treated HepG2 cells. A: HepG2 cells were transfected with control siRNA or G0S2 siRNA, and G0S2 expression was measured by Western blotting. At least three independent experiments were conducted. Data are presented as means ± SE. f P

Techniques Used: Expressing, Transfection, Western Blot

46) Product Images from "Mitochondria Dysfunction in Aging and Metabolic Diseases: The E3 ligase MARCH5 is a PPARγ target gene that regulates mitochondria and metabolism in adipocytes"

Article Title: Mitochondria Dysfunction in Aging and Metabolic Diseases: The E3 ligase MARCH5 is a PPARγ target gene that regulates mitochondria and metabolism in adipocytes

Journal: American Journal of Physiology - Endocrinology and Metabolism

doi: 10.1152/ajpendo.00394.2018

Peroxisome proliferator-activated receptor-γ (PPARγ) and mitochondrial protein expression, including membrane-associated RING finger (C3HC4) 5 (MARCH5) are increased during adipocyte differentiation. A : Western blots and densitometric quantification of MARCH5 and mitofusin2 (Mfn2) from wild-type (WT) and MARCH5 knockout (KO) mouse embryonic fibroblasts (MEFs), adjusted for loading control (14-3-3). B and C : Western blot and densitometry analyses of adipogenic and mitochondrial proteins from predifferentiated through day 10- postdifferentiated 3T3-L1 adipocytes. Values are means ± SE; n = 60,000 cells/well. * P
Figure Legend Snippet: Peroxisome proliferator-activated receptor-γ (PPARγ) and mitochondrial protein expression, including membrane-associated RING finger (C3HC4) 5 (MARCH5) are increased during adipocyte differentiation. A : Western blots and densitometric quantification of MARCH5 and mitofusin2 (Mfn2) from wild-type (WT) and MARCH5 knockout (KO) mouse embryonic fibroblasts (MEFs), adjusted for loading control (14-3-3). B and C : Western blot and densitometry analyses of adipogenic and mitochondrial proteins from predifferentiated through day 10- postdifferentiated 3T3-L1 adipocytes. Values are means ± SE; n = 60,000 cells/well. * P

Techniques Used: Expressing, Western Blot, Knock-Out

Membrane-associated RING finger (C3HC4) 5 (MARCH5) is highly expressed in adipose tissue and is regulated by peroxisome proliferator-activated receptor-γ (PPARγ). A : dot plot depicting genes significantly (5% false discovery rate) positively correlated (bicorrelation r > 0.4) with PPARγ expression in white adipose tissue (WAT) of Hybrid Mouse Diversity Panel (HMDP) strains; red dots, previously validated PPARγ target genes; blue dot, March5 . Hibadh , 3-hydroxyisobutyrate dehydrogenase; OsbpL11 , oxysterol-binding protein-like 11; Pex13 , peroxisomal biogenesis factor 13. B : pathway enrichment analysis of genes correlated (bicorrelation r > 0.5) with PPARγ in WAT from strains of the HMDP. C : Western blots for MARCH5 and 14-3-3 (loading control) in WAT and brown adipose tissue (BAT) from C57BL/6J mice. D : gene expression analysis for March5 mRNA in various tissues of C57BL/6J mice. Hrt, heart; Quad, quadriceps; Sol, soleus; Liv, liver. Values are means ± SE; n = 5. E : chromatin immunoprecipitation-sequencing analysis of PPARγ, retinoic acid receptor-α (RXRα), and CCAAT enhancer-binding protein-α (C/EBPα) at the March5 locus in 3T3-L1 adipocytes. F : photomicrographs of predifferentiated (Pre-Diff) and differentiated ( day 7 Diff) primary adipocytes isolated from WT (PPARγ floxed) and PPARγ knockout [PPARγ floxed + aP2-cre (PPARγ-KO)] C57BL/6J mice. G : gene expression analysis for March5 , normalized to cyclophilin A ( Ppia ), in cells shown in photomicrographs in F . PPg-KO, PPARγ-KO. Values [arbitrary units (AU)] are means ± SE. * P
Figure Legend Snippet: Membrane-associated RING finger (C3HC4) 5 (MARCH5) is highly expressed in adipose tissue and is regulated by peroxisome proliferator-activated receptor-γ (PPARγ). A : dot plot depicting genes significantly (5% false discovery rate) positively correlated (bicorrelation r > 0.4) with PPARγ expression in white adipose tissue (WAT) of Hybrid Mouse Diversity Panel (HMDP) strains; red dots, previously validated PPARγ target genes; blue dot, March5 . Hibadh , 3-hydroxyisobutyrate dehydrogenase; OsbpL11 , oxysterol-binding protein-like 11; Pex13 , peroxisomal biogenesis factor 13. B : pathway enrichment analysis of genes correlated (bicorrelation r > 0.5) with PPARγ in WAT from strains of the HMDP. C : Western blots for MARCH5 and 14-3-3 (loading control) in WAT and brown adipose tissue (BAT) from C57BL/6J mice. D : gene expression analysis for March5 mRNA in various tissues of C57BL/6J mice. Hrt, heart; Quad, quadriceps; Sol, soleus; Liv, liver. Values are means ± SE; n = 5. E : chromatin immunoprecipitation-sequencing analysis of PPARγ, retinoic acid receptor-α (RXRα), and CCAAT enhancer-binding protein-α (C/EBPα) at the March5 locus in 3T3-L1 adipocytes. F : photomicrographs of predifferentiated (Pre-Diff) and differentiated ( day 7 Diff) primary adipocytes isolated from WT (PPARγ floxed) and PPARγ knockout [PPARγ floxed + aP2-cre (PPARγ-KO)] C57BL/6J mice. G : gene expression analysis for March5 , normalized to cyclophilin A ( Ppia ), in cells shown in photomicrographs in F . PPg-KO, PPARγ-KO. Values [arbitrary units (AU)] are means ± SE. * P

Techniques Used: Expressing, Binding Assay, Western Blot, Mouse Assay, ChIP-sequencing, Isolation, Knock-Out

47) Product Images from "Peroxisome Proliferator Activated Receptor gamma (PPARγ) Agonist Rosiglitazone Ameliorate Airway Inflammation by Inhibiting Toll-Like Receptor 2 (TLR2)/Nod-Like Receptor with Pyrin Domain Containing 3 (NLRP3) Inflammatory Corpuscle Activation in Asthmatic Mice"

Article Title: Peroxisome Proliferator Activated Receptor gamma (PPARγ) Agonist Rosiglitazone Ameliorate Airway Inflammation by Inhibiting Toll-Like Receptor 2 (TLR2)/Nod-Like Receptor with Pyrin Domain Containing 3 (NLRP3) Inflammatory Corpuscle Activation in Asthmatic Mice

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

doi: 10.12659/MSM.910766

Expression of toll-like receptor 2 (TLR2) and peroxisome proliferator activated receptor agonist (PPARγ) proteins were significantly decreased after rosiglitazone treatment. ( A ) Expression of TLR2 and PPARγ proteins by western blot analysis. ( B ) Quantitative analysis of protein test strips. Data are represented as the mean ±SD. * P
Figure Legend Snippet: Expression of toll-like receptor 2 (TLR2) and peroxisome proliferator activated receptor agonist (PPARγ) proteins were significantly decreased after rosiglitazone treatment. ( A ) Expression of TLR2 and PPARγ proteins by western blot analysis. ( B ) Quantitative analysis of protein test strips. Data are represented as the mean ±SD. * P

Techniques Used: Expressing, Western Blot

48) Product Images from "Pituitary Adenylate Cyclase Activating Peptide (PACAP) Participates in Adipogenesis by Activating ERK Signaling Pathway"

Article Title: Pituitary Adenylate Cyclase Activating Peptide (PACAP) Participates in Adipogenesis by Activating ERK Signaling Pathway

Journal: PLoS ONE

doi: 10.1371/journal.pone.0072607

Expression of C/EBPα/β, PPARγ and aquaporin 7 (AQP7) during differentiation of 3T3-L1 cells by XDI and/or PDI cocktail. A. Quantification of mRNA expression levels of crucial transcription factors, C/EBPα/β, PPARγ and AQP7 a PPARγ target, at the indicated times of differentiation in mouse 3T3-L1 adipocytes. Cells were induced to differentiate with XDI (shaded) or PDI (black) hormonal cocktail. C/EBPα, PPARγ and AQP7 mRNA expression levels were measured by qPCR during TD till day 9. cEBPβ mRNA expression levels were measured during MCE. Data were analyzed using repeated measure of ANOVA and by Tukey’s comparison tests. *p
Figure Legend Snippet: Expression of C/EBPα/β, PPARγ and aquaporin 7 (AQP7) during differentiation of 3T3-L1 cells by XDI and/or PDI cocktail. A. Quantification of mRNA expression levels of crucial transcription factors, C/EBPα/β, PPARγ and AQP7 a PPARγ target, at the indicated times of differentiation in mouse 3T3-L1 adipocytes. Cells were induced to differentiate with XDI (shaded) or PDI (black) hormonal cocktail. C/EBPα, PPARγ and AQP7 mRNA expression levels were measured by qPCR during TD till day 9. cEBPβ mRNA expression levels were measured during MCE. Data were analyzed using repeated measure of ANOVA and by Tukey’s comparison tests. *p

Techniques Used: Expressing, Real-time Polymerase Chain Reaction

49) Product Images from "Lipoatrophy and severe metabolic disturbance in mice with fat-specific deletion of PPAR?"

Article Title: Lipoatrophy and severe metabolic disturbance in mice with fat-specific deletion of PPAR?

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1314863110

Delayed hair coat formation, arrested mammary gland development and high bone mass in PPARγ FKO mice. ( A ) H E ( i ) and Oil Red O ( ii ) staining sections of skin from 8-d-old male control and PPARγ FKO pups. ( B ) Whole-mount analysis
Figure Legend Snippet: Delayed hair coat formation, arrested mammary gland development and high bone mass in PPARγ FKO mice. ( A ) H E ( i ) and Oil Red O ( ii ) staining sections of skin from 8-d-old male control and PPARγ FKO pups. ( B ) Whole-mount analysis

Techniques Used: Mouse Assay, Staining

Loss of WAT, pale liver, and hyperlipidemia in 8-d-old PPARγ FKO pups. ( A ) Exposed ventral view of the 8-d-old male control and PPARγ FKO mice, illustrating the pale liver (yellow arrows) and reduction of IWAT depots (white arrows) in
Figure Legend Snippet: Loss of WAT, pale liver, and hyperlipidemia in 8-d-old PPARγ FKO pups. ( A ) Exposed ventral view of the 8-d-old male control and PPARγ FKO mice, illustrating the pale liver (yellow arrows) and reduction of IWAT depots (white arrows) in

Techniques Used: Mouse Assay

Severe lipoatrophy in 3-mo-old PPARγ FKO mice. ( A – E ) Gross morphology of GWAT (white arrow) and liver (yellow arrow) ( A ), interscapular fat ( B ), IWAT ( C ), mesenteric WAT ( D ), and perirenal WAT ( E ) from 3-mo-old male control and PPARγ
Figure Legend Snippet: Severe lipoatrophy in 3-mo-old PPARγ FKO mice. ( A – E ) Gross morphology of GWAT (white arrow) and liver (yellow arrow) ( A ), interscapular fat ( B ), IWAT ( C ), mesenteric WAT ( D ), and perirenal WAT ( E ) from 3-mo-old male control and PPARγ

Techniques Used: Mouse Assay

Specific deletion of PPARγ in adipose tissues. ( A ) PPARγ mRNA level of 3-mo-old control and PPARγ FHet mice in BAT, IWAT, GWAT, liver, muscle, and brain. Values are mean ± SEM ( n = 4–5). PPARγ primers are
Figure Legend Snippet: Specific deletion of PPARγ in adipose tissues. ( A ) PPARγ mRNA level of 3-mo-old control and PPARγ FHet mice in BAT, IWAT, GWAT, liver, muscle, and brain. Values are mean ± SEM ( n = 4–5). PPARγ primers are

Techniques Used: Mouse Assay

Ablation of BAT in 1-d-old PPARγ FKO mice. ( A ) Gross morphology of interscapular BAT (arrows) from 1-d-old male control and PPARγ FKO pups. ( B ) H E staining of transverse sections from the interscapular region of 1-d-old male PPARγ
Figure Legend Snippet: Ablation of BAT in 1-d-old PPARγ FKO mice. ( A ) Gross morphology of interscapular BAT (arrows) from 1-d-old male control and PPARγ FKO pups. ( B ) H E staining of transverse sections from the interscapular region of 1-d-old male PPARγ

Techniques Used: Mouse Assay, Staining

Massive fatty livers and extreme insulin resistance in 3-mo-old PPARγ FKO mice. ( A ) Liver weights from male or female 3-mo-old control and PPARγ FKO mice. ( B ) H E staining of livers from 3-mo-old female control and PPARγ
Figure Legend Snippet: Massive fatty livers and extreme insulin resistance in 3-mo-old PPARγ FKO mice. ( A ) Liver weights from male or female 3-mo-old control and PPARγ FKO mice. ( B ) H E staining of livers from 3-mo-old female control and PPARγ

Techniques Used: Mouse Assay, Staining

50) Product Images from "TLE3 is a dual function transcriptional coregulator of adipogenesis"

Article Title: TLE3 is a dual function transcriptional coregulator of adipogenesis

Journal: Cell metabolism

doi: 10.1016/j.cmet.2011.02.014

TLE3 is a PPARγ target gene (A) Realtime PCR analysis of TLE mRNA expression in PPARγ2-expressing 10T1/2 preadipocytes treated with 100 nM GW7845 for 2 d (left) and 3T3-L1 preadipocytes treated with DMI + 20 nM GW for 2 d (right). (B) Induction of TLE3 and aP2 mRNA by PPARγ agonist in white and brown adipose tissue in vivo .
Figure Legend Snippet: TLE3 is a PPARγ target gene (A) Realtime PCR analysis of TLE mRNA expression in PPARγ2-expressing 10T1/2 preadipocytes treated with 100 nM GW7845 for 2 d (left) and 3T3-L1 preadipocytes treated with DMI + 20 nM GW for 2 d (right). (B) Induction of TLE3 and aP2 mRNA by PPARγ agonist in white and brown adipose tissue in vivo .

Techniques Used: Polymerase Chain Reaction, Expressing, In Vivo

TLE3 coactivates PPARγ-dependent gene expression .
Figure Legend Snippet: TLE3 coactivates PPARγ-dependent gene expression .

Techniques Used: Expressing

Overlapping transcriptional profiles of PPARγ and TLE3 regulated genes .
Figure Legend Snippet: Overlapping transcriptional profiles of PPARγ and TLE3 regulated genes .

Techniques Used:

51) Product Images from "Activation of PPARγ Ameliorates Spatial Cognitive Deficits through Restoring Expression of AMPA Receptors in Seipin Knock-Out Mice"

Article Title: Activation of PPARγ Ameliorates Spatial Cognitive Deficits through Restoring Expression of AMPA Receptors in Seipin Knock-Out Mice

Journal: The Journal of Neuroscience

doi: 10.1523/JNEUROSCI.3280-15.2016

Neuronal seipin deficiency reduces hippocampal PPARγ level. A , Representative images of seipin immunostaining in hippocampal CA1 region of control mice and seipin -nKO mice. str-R, Stratum radiatum; str-P, stratum pyramidale; str-O, stratum oriens. Arrows indicate seipin-immunopositive cells. Scale bars, 50 μm. B , Bars show the levels of hippocampal PPARγ in seipin -sKO mice and WT mice, seipin -nKO mice and control mice, or seipin -aKO mice and a-control mice treated with vehicle or rosi for 3 d. The densitometric values for Western blots of protein are normalized to the amounts of GAPDH and normalized again by control values. ** p
Figure Legend Snippet: Neuronal seipin deficiency reduces hippocampal PPARγ level. A , Representative images of seipin immunostaining in hippocampal CA1 region of control mice and seipin -nKO mice. str-R, Stratum radiatum; str-P, stratum pyramidale; str-O, stratum oriens. Arrows indicate seipin-immunopositive cells. Scale bars, 50 μm. B , Bars show the levels of hippocampal PPARγ in seipin -sKO mice and WT mice, seipin -nKO mice and control mice, or seipin -aKO mice and a-control mice treated with vehicle or rosi for 3 d. The densitometric values for Western blots of protein are normalized to the amounts of GAPDH and normalized again by control values. ** p

Techniques Used: Immunostaining, Mouse Assay, Western Blot

52) Product Images from "Targeting FoxO1 with AS1842856 Suppresses Adipogenesis"

Article Title: Targeting FoxO1 with AS1842856 Suppresses Adipogenesis

Journal: Cell Cycle

doi: 10.4161/15384101.2014.965977

AS1842856 suppressed PPARγ and mitochondrial protein expression. ( A ) Western blots showing the effect of AS1842856 on PPARγ, adiponectin, mitochondrial proteins C1 and C3. β-actin was probed as the loading control. DI, differentiation induction; AS, AS1842856. ( B–D ) Densitometric analysis of western blot images with NIH ImageJ software; n = 3−5. * P
Figure Legend Snippet: AS1842856 suppressed PPARγ and mitochondrial protein expression. ( A ) Western blots showing the effect of AS1842856 on PPARγ, adiponectin, mitochondrial proteins C1 and C3. β-actin was probed as the loading control. DI, differentiation induction; AS, AS1842856. ( B–D ) Densitometric analysis of western blot images with NIH ImageJ software; n = 3−5. * P

Techniques Used: Expressing, Western Blot, Software

Kinetics of FoxO1-regulated protein expression during adipogenesis. ( A ) Western blots showing the expression of PPARγ, adiponectin, mitochondrial proteins C1 and C3. β-actin was probed as the loading control. ( B ) Densitometric analysis of western blot images for PPARγ and adiponectin with NIH ImageJ software. ( C ) Densitometric analysis of protein gel blot images for C1 and C3 with NIH ImageJ software. n = 3−5. * P
Figure Legend Snippet: Kinetics of FoxO1-regulated protein expression during adipogenesis. ( A ) Western blots showing the expression of PPARγ, adiponectin, mitochondrial proteins C1 and C3. β-actin was probed as the loading control. ( B ) Densitometric analysis of western blot images for PPARγ and adiponectin with NIH ImageJ software. ( C ) Densitometric analysis of protein gel blot images for C1 and C3 with NIH ImageJ software. n = 3−5. * P

Techniques Used: Expressing, Western Blot, Software

53) Product Images from "Isobavachalcone from Angelica keiskei Inhibits Adipogenesis and Prevents Lipid Accumulation"

Article Title: Isobavachalcone from Angelica keiskei Inhibits Adipogenesis and Prevents Lipid Accumulation

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19061693

Effect of IBC on autophagic flux during adipocyte differentiation. ( A ) 3T3-L1 adipocytes were differentiated in the presence or absence of 40 μM of IBC. Differentiating adipocytes were harvested at the indicated period and lysed for Western blotting analysis to determine levels of LC3B and SQSTM1/p62; ( B ) Preadipocytes were transfected with GFP-LC3 plasmid and differentiated with MDI in the presence of IBC or CQ (10 μM) during D0–D2, followed by additional treatment with differentiation medium for 2 days (D4). Cells were fixed and stained with DAPI (nuclei, blue-colored). Intracellular GFP-LC3 puncta (green-colored) were visualized with a confocal laser microscope. Scale bar = 10 μm; ( C ) Differentiating adipocytes supplemented with IBC or CQ during D0–D2 were harvested and protein levels of PPARγ and SQSTM1/p62 were analyzed on D8. Scale bar = 100 μm; and ( D ) On D2, differentiating adipocytes supplemented with IBC or CQ were harvested and subjected to qPCR to analyze gene expression levels of BECN1 , Atg5 , and Atg7 . Data are presented as means ± SD of triplicate experiments. * p
Figure Legend Snippet: Effect of IBC on autophagic flux during adipocyte differentiation. ( A ) 3T3-L1 adipocytes were differentiated in the presence or absence of 40 μM of IBC. Differentiating adipocytes were harvested at the indicated period and lysed for Western blotting analysis to determine levels of LC3B and SQSTM1/p62; ( B ) Preadipocytes were transfected with GFP-LC3 plasmid and differentiated with MDI in the presence of IBC or CQ (10 μM) during D0–D2, followed by additional treatment with differentiation medium for 2 days (D4). Cells were fixed and stained with DAPI (nuclei, blue-colored). Intracellular GFP-LC3 puncta (green-colored) were visualized with a confocal laser microscope. Scale bar = 10 μm; ( C ) Differentiating adipocytes supplemented with IBC or CQ during D0–D2 were harvested and protein levels of PPARγ and SQSTM1/p62 were analyzed on D8. Scale bar = 100 μm; and ( D ) On D2, differentiating adipocytes supplemented with IBC or CQ were harvested and subjected to qPCR to analyze gene expression levels of BECN1 , Atg5 , and Atg7 . Data are presented as means ± SD of triplicate experiments. * p

Techniques Used: Western Blot, Transfection, Plasmid Preparation, Staining, Microscopy, Real-time Polymerase Chain Reaction, Expressing

54) Product Images from "PPARγ agonists promote differentiation of cancer stem cells by restraining YAP transcriptional activity"

Article Title: PPARγ agonists promote differentiation of cancer stem cells by restraining YAP transcriptional activity

Journal: Oncotarget

doi: 10.18632/oncotarget.11273

A CTGF-PPARγ-TIMP3 signature correlates with clinical outcome in osteosarcoma Kaplan-Meier survival curves for CTGF, PPARγ and TIMP3 illustrating that higher CTGF, and PPARγ and low TIMP3 expression correlates with worse outcomes, and the reverse pattern of expression correlates with better outcome in two independent datasets. Distribution of patients for this data set has been previously published. Survival probability and P values calculated using Kaplan-Meier and Cox proportional hazards methods. DFS = Disease-free survival in Kelly, OS = Overall survival in TCGA.
Figure Legend Snippet: A CTGF-PPARγ-TIMP3 signature correlates with clinical outcome in osteosarcoma Kaplan-Meier survival curves for CTGF, PPARγ and TIMP3 illustrating that higher CTGF, and PPARγ and low TIMP3 expression correlates with worse outcomes, and the reverse pattern of expression correlates with better outcome in two independent datasets. Distribution of patients for this data set has been previously published. Survival probability and P values calculated using Kaplan-Meier and Cox proportional hazards methods. DFS = Disease-free survival in Kelly, OS = Overall survival in TCGA.

Techniques Used: Expressing

Rosiglitazone decreases YAP nuclear localization and YAP-dependent transcription in osteosarcoma cells A. Expression of canonical YAP target genes in mOS-482 cells treated with 100 uM rosiglitazone for 48 hours. B. Western blot of YAP and phospho-YAP C. Immunofluorescence using Sox2, YAP and RNA Pol II antibody on mOS-482 cells treated with Rosi or adipogenic media for 48 hours. Images were taken using a Leica DM5500 immunofluorescence microscope at 63x magnification. D. YAP localization by TZD is dependent on PPARγ. Quantification of YAP immunostaining in CAS9 (control) and PPARγ knockout cells treatedwith 50 μM Rosi for 24 hours. Images were taken using a Leica DM5500 immunofluorescence microscope at 63x magnification. Ten fields in each condition were counted and average percentage of cells showing exclusively nuclear (N), exclusively cytoplasmic (C), or both nuclear and cytoplasmic (N+C) is shown in histogram. *p
Figure Legend Snippet: Rosiglitazone decreases YAP nuclear localization and YAP-dependent transcription in osteosarcoma cells A. Expression of canonical YAP target genes in mOS-482 cells treated with 100 uM rosiglitazone for 48 hours. B. Western blot of YAP and phospho-YAP C. Immunofluorescence using Sox2, YAP and RNA Pol II antibody on mOS-482 cells treated with Rosi or adipogenic media for 48 hours. Images were taken using a Leica DM5500 immunofluorescence microscope at 63x magnification. D. YAP localization by TZD is dependent on PPARγ. Quantification of YAP immunostaining in CAS9 (control) and PPARγ knockout cells treatedwith 50 μM Rosi for 24 hours. Images were taken using a Leica DM5500 immunofluorescence microscope at 63x magnification. Ten fields in each condition were counted and average percentage of cells showing exclusively nuclear (N), exclusively cytoplasmic (C), or both nuclear and cytoplasmic (N+C) is shown in histogram. *p

Techniques Used: Expressing, Western Blot, Immunofluorescence, Microscopy, Immunostaining, Knock-Out

TZD treatment induces adipogenesis in osteosarcoma cells in part through PPARγ activation A. Oil Red-O lipid stain of mOS-cells grown in adipogenic media or Rosiglitazone (Rosi) 10uM or Pioglitazone (Pio) 10 uM for 3 days. Mag 40X. Right panel - Relative fold change in mRNA expression of FABP4 measured by qRT-PCR relative to actin as a control. B. mOS control, Cas9-expressing or Cas9-PPARγ knockout cells were treated with increasing concentrations of Rosi, as indicated and cell number was determined after 48 hours. Right Panel - Western blot confirming PPARγ deletion in mOS cells expressing PPARγ specific guide RNA.
Figure Legend Snippet: TZD treatment induces adipogenesis in osteosarcoma cells in part through PPARγ activation A. Oil Red-O lipid stain of mOS-cells grown in adipogenic media or Rosiglitazone (Rosi) 10uM or Pioglitazone (Pio) 10 uM for 3 days. Mag 40X. Right panel - Relative fold change in mRNA expression of FABP4 measured by qRT-PCR relative to actin as a control. B. mOS control, Cas9-expressing or Cas9-PPARγ knockout cells were treated with increasing concentrations of Rosi, as indicated and cell number was determined after 48 hours. Right Panel - Western blot confirming PPARγ deletion in mOS cells expressing PPARγ specific guide RNA.

Techniques Used: Activation Assay, Staining, Expressing, Quantitative RT-PCR, Knock-Out, Western Blot

TZDs target the cancer stem cell population of osteosarcomas mOS-482 cells were fractionated into Sca-1 High and Sca-1 Low fractions by fluorescence activated cell sorting (FACS). The two fractions were treated with 50 or 100 uM Rosiglitazone (Rosi), and A. , cell proliferation and B. , expression of adipocyte-marker FABP4 by qRT-PCR, were measured. C. Relative fold change in mRNA expression of PPARγ measured by qRT-PCR relative to actin as a control. D. Flow cytometric analysis of membrane Sca-1 expression of phycoerythrin-labeled (Sca-1-PE) mOS-482 cells before and after treatment with 100 μM Rosi for 72 hours. The histogram shows mean fluorescence intensity of the indicated cells. Y axis is maximum mean fluorescence intensity. X axis is IgG-phycoerythrin stained cells (-phycoerythrin-conjugated - anti Sca-antibody).
Figure Legend Snippet: TZDs target the cancer stem cell population of osteosarcomas mOS-482 cells were fractionated into Sca-1 High and Sca-1 Low fractions by fluorescence activated cell sorting (FACS). The two fractions were treated with 50 or 100 uM Rosiglitazone (Rosi), and A. , cell proliferation and B. , expression of adipocyte-marker FABP4 by qRT-PCR, were measured. C. Relative fold change in mRNA expression of PPARγ measured by qRT-PCR relative to actin as a control. D. Flow cytometric analysis of membrane Sca-1 expression of phycoerythrin-labeled (Sca-1-PE) mOS-482 cells before and after treatment with 100 μM Rosi for 72 hours. The histogram shows mean fluorescence intensity of the indicated cells. Y axis is maximum mean fluorescence intensity. X axis is IgG-phycoerythrin stained cells (-phycoerythrin-conjugated - anti Sca-antibody).

Techniques Used: Fluorescence, FACS, Expressing, Marker, Quantitative RT-PCR, Flow Cytometry, Labeling, Staining

55) Product Images from "Hepatocyte-specific Nrf2 deficiency mitigates high-fat diet-induced hepatic steatosis: Involvement of reduced PPARγ expression"

Article Title: Hepatocyte-specific Nrf2 deficiency mitigates high-fat diet-induced hepatic steatosis: Involvement of reduced PPARγ expression

Journal: Redox Biology

doi: 10.1016/j.redox.2019.101412

Nrf2 (L)-KO hepatocytes exhibit decreased response to PPARγ agonists. (A–D) Primary hepatocytes isolated from Nrf2 -LoxP and Nrf2 (L)-KO mice fed with CD were cultured for 6 h in normal media and then exposed to rosiglitazone (ROSI, Veh, 1, 10 μM) or pioglitazone (PIOG, Veh, 0.25, 0.5 μM) for 24 h. (A–D) The mRNA expression (A and C) and representative images of immunoblots of PPARγ (B and D) in the cells treated with ROSI (A and B) or PIGO (C and D). * p
Figure Legend Snippet: Nrf2 (L)-KO hepatocytes exhibit decreased response to PPARγ agonists. (A–D) Primary hepatocytes isolated from Nrf2 -LoxP and Nrf2 (L)-KO mice fed with CD were cultured for 6 h in normal media and then exposed to rosiglitazone (ROSI, Veh, 1, 10 μM) or pioglitazone (PIOG, Veh, 0.25, 0.5 μM) for 24 h. (A–D) The mRNA expression (A and C) and representative images of immunoblots of PPARγ (B and D) in the cells treated with ROSI (A and B) or PIGO (C and D). * p

Techniques Used: Isolation, Mouse Assay, Cell Culture, Expressing, Western Blot

Schematic illustration of the proposed role of N RF 2 in hepatocytes in HFD-induced NAFLD. Following a prolonged HFD exposure, the two genotypes of mice displayed different liver mass and TG accumulation in the liver. A possible mechanism is that NRF2 in hepatocyte is responsible for the expression of PPARγ to positively regulate lipid accumulation. Ablation of Nrf2 in hepatocytes results in down-regulation of PPARγ expression and subsequent reduction of lipid accumulation in the cells. The key findings in the present study demonstrate an essential role of NRF2 in the initiation of NAFLD via PPARγ expression.
Figure Legend Snippet: Schematic illustration of the proposed role of N RF 2 in hepatocytes in HFD-induced NAFLD. Following a prolonged HFD exposure, the two genotypes of mice displayed different liver mass and TG accumulation in the liver. A possible mechanism is that NRF2 in hepatocyte is responsible for the expression of PPARγ to positively regulate lipid accumulation. Ablation of Nrf2 in hepatocytes results in down-regulation of PPARγ expression and subsequent reduction of lipid accumulation in the cells. The key findings in the present study demonstrate an essential role of NRF2 in the initiation of NAFLD via PPARγ expression.

Techniques Used: Mouse Assay, Expressing

Nrf2 (L)-KO hepatocytes show reduced activation of PPARγ in response to palmitate challenge. (A–B) The mRNA levels of Pparg1 and Pparg2 . Isolated primary hepatocytes from Nrf2 -LoxP and Nrf2 (L)-KO mice were challenged with 0.5 mM palmitate for indicated time. * p
Figure Legend Snippet: Nrf2 (L)-KO hepatocytes show reduced activation of PPARγ in response to palmitate challenge. (A–B) The mRNA levels of Pparg1 and Pparg2 . Isolated primary hepatocytes from Nrf2 -LoxP and Nrf2 (L)-KO mice were challenged with 0.5 mM palmitate for indicated time. * p

Techniques Used: Activation Assay, Isolation, Mouse Assay

The reduced expression of PPARγ-related genes in Nrf2 (L)-KO hepatocytes is partially reversed by PPARγ overexpression. Hepatocytes isolated from Nrf2 -LoxP and Nrf2 (L)-KO mice were transfected with Pparg1 and Pparg2 plasmids. (A) mRNA levels of Pparg1 and Pparg2 in the cells. * p
Figure Legend Snippet: The reduced expression of PPARγ-related genes in Nrf2 (L)-KO hepatocytes is partially reversed by PPARγ overexpression. Hepatocytes isolated from Nrf2 -LoxP and Nrf2 (L)-KO mice were transfected with Pparg1 and Pparg2 plasmids. (A) mRNA levels of Pparg1 and Pparg2 in the cells. * p

Techniques Used: Expressing, Over Expression, Isolation, Mouse Assay, Transfection

56) Product Images from "p38 MAPK-inhibited dendritic cells induce superior antitumor immune responses and overcome regulatory T cell-mediated immunosuppression"

Article Title: p38 MAPK-inhibited dendritic cells induce superior antitumor immune responses and overcome regulatory T cell-mediated immunosuppression

Journal: Nature communications

doi: 10.1038/ncomms5229

Downregulation of PPARγ expression/activity in mSBDCs contributes to the p50-mediated increased expression of surface OX40L. ( a ) Western blot analysis of the PPARγ, pC/EBPβ and C/EBPβ in mDCs and mSBDCs treated with or without p38 inhibitor SB202190, PPARγ inhibitor GW9662, and/or PPARγ activator RGZ. ( b ) The protein level of PPARγ, pC/EBPβ and C/EBPβ in mDCs treated with C/EBPβ-specific siRNA. ( c ) RT-PCR analysis of Ap2 mRNA expression in mDCs treated with or without p38 inhibitor SB202190, PPARγ inhibitor GW9662, PPARγ activator RGZ, and/or p38 activator anisomycin. Data shown (n = 3) were normalized to the Gapdh gene. Error bars represent s.d. ( d ) Western blot analysis of the cytoplasmic and nuclear localization of p50 molecule in treated mDCs and mSBDCs. ( e ) Coimmunoprecipitation of endogenous p50 and PPARγ. mDCs and mSBDCs lysates were immunoprecipitated with PPARγ antibody or isotype control IgG and immunoblotted with p50 antibody. ( f ) Surface expression of OX40L on treated mDCs and mSBDCs generated from wild-type and p50 −/− mice analyzed by FACS. Numbers showed the percentage of OX40L-positive DCs gated on CD11c + cells. ( g ) Summarized data (n = 4) from ( f ). Error bars represent s.d. Representative data from one of two performed experiments are shown. P values were calculated with Student’s t -test.
Figure Legend Snippet: Downregulation of PPARγ expression/activity in mSBDCs contributes to the p50-mediated increased expression of surface OX40L. ( a ) Western blot analysis of the PPARγ, pC/EBPβ and C/EBPβ in mDCs and mSBDCs treated with or without p38 inhibitor SB202190, PPARγ inhibitor GW9662, and/or PPARγ activator RGZ. ( b ) The protein level of PPARγ, pC/EBPβ and C/EBPβ in mDCs treated with C/EBPβ-specific siRNA. ( c ) RT-PCR analysis of Ap2 mRNA expression in mDCs treated with or without p38 inhibitor SB202190, PPARγ inhibitor GW9662, PPARγ activator RGZ, and/or p38 activator anisomycin. Data shown (n = 3) were normalized to the Gapdh gene. Error bars represent s.d. ( d ) Western blot analysis of the cytoplasmic and nuclear localization of p50 molecule in treated mDCs and mSBDCs. ( e ) Coimmunoprecipitation of endogenous p50 and PPARγ. mDCs and mSBDCs lysates were immunoprecipitated with PPARγ antibody or isotype control IgG and immunoblotted with p50 antibody. ( f ) Surface expression of OX40L on treated mDCs and mSBDCs generated from wild-type and p50 −/− mice analyzed by FACS. Numbers showed the percentage of OX40L-positive DCs gated on CD11c + cells. ( g ) Summarized data (n = 4) from ( f ). Error bars represent s.d. Representative data from one of two performed experiments are shown. P values were calculated with Student’s t -test.

Techniques Used: Expressing, Activity Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction, Immunoprecipitation, Generated, Mouse Assay, FACS

The role of p38 MAPK signaling in the regulation of DC immunogenicity. p38 MAPK phosphorylates C/EBPβ and pC/EBPβ induces the production of PPARγ, PPARγ can be activated by its ligand or agonist, which can then repress the transactivation of p50 by physical interaction with p50 to prevent its binding to the response elements such as Ox40l promoter. p38 MAPK inhibitor, such as SB202190, could abrogate this self-limitation break and result in a robust p50 nuclear translocation. p50, after forming transcriptionally active complexes with RelB, induces the strikingly increased surface OX40L expression, which is crucial to activation of Teff and inhibition of Treg.
Figure Legend Snippet: The role of p38 MAPK signaling in the regulation of DC immunogenicity. p38 MAPK phosphorylates C/EBPβ and pC/EBPβ induces the production of PPARγ, PPARγ can be activated by its ligand or agonist, which can then repress the transactivation of p50 by physical interaction with p50 to prevent its binding to the response elements such as Ox40l promoter. p38 MAPK inhibitor, such as SB202190, could abrogate this self-limitation break and result in a robust p50 nuclear translocation. p50, after forming transcriptionally active complexes with RelB, induces the strikingly increased surface OX40L expression, which is crucial to activation of Teff and inhibition of Treg.

Techniques Used: Binding Assay, Translocation Assay, Expressing, Activation Assay, Inhibition

57) Product Images from "Nicotinamide Promotes Adipogenesis in Umbilical Cord-Derived Mesenchymal Stem Cells and Is Associated with Neonatal Adiposity: The Healthy Start BabyBUMP Project"

Article Title: Nicotinamide Promotes Adipogenesis in Umbilical Cord-Derived Mesenchymal Stem Cells and Is Associated with Neonatal Adiposity: The Healthy Start BabyBUMP Project

Journal: PLoS ONE

doi: 10.1371/journal.pone.0159575

Scatter plots depicting the correlation between neonatal adiposity and the percent change of PPARγ (A) and FABP4 (B) protein content at day 21 between vehicle-control and NAM only conditions (N = 46).
Figure Legend Snippet: Scatter plots depicting the correlation between neonatal adiposity and the percent change of PPARγ (A) and FABP4 (B) protein content at day 21 between vehicle-control and NAM only conditions (N = 46).

Techniques Used:

NAM increases PPARγ, FABP4, and lipid content in MSC-derived adipocytes. MSCs were incubated with standard differentiation media for 21 days +/-NAM (3mM) as described in Methods. Protein content at day 21 was measured by ICE assay in all treatment conditions (A and B) (N = 46). *p
Figure Legend Snippet: NAM increases PPARγ, FABP4, and lipid content in MSC-derived adipocytes. MSCs were incubated with standard differentiation media for 21 days +/-NAM (3mM) as described in Methods. Protein content at day 21 was measured by ICE assay in all treatment conditions (A and B) (N = 46). *p

Techniques Used: Derivative Assay, Incubation

Effects of NAM (3mM) incubation during adipocyte cell differentiation in human MSC. Cells were harvested at day 9 of differentiation and mRNA expression of PPARγ (A), SIRT1 enzyme activity (B), and acetylation of SIRT1 protein targets, PPARγ (C) and β-catenin (D), and mRNA expression of NAMPT (E) in the vehicle-control and NAM only conditions as described in Methods. N = 9 per group. *p
Figure Legend Snippet: Effects of NAM (3mM) incubation during adipocyte cell differentiation in human MSC. Cells were harvested at day 9 of differentiation and mRNA expression of PPARγ (A), SIRT1 enzyme activity (B), and acetylation of SIRT1 protein targets, PPARγ (C) and β-catenin (D), and mRNA expression of NAMPT (E) in the vehicle-control and NAM only conditions as described in Methods. N = 9 per group. *p

Techniques Used: Incubation, Cell Differentiation, Expressing, Activity Assay

58) Product Images from "Licochalcone E protects against carbon tetrachloride-induced liver toxicity by activating peroxisome proliferator-activated receptor gamma"

Article Title: Licochalcone E protects against carbon tetrachloride-induced liver toxicity by activating peroxisome proliferator-activated receptor gamma

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2017.7268

Effect of LCE on PPARγ and NF-κB protein levels in mice liver tissue. (A) Measurement of PPARγ and NF-κB protein levels in mice liver were analyzed by western blot. Expression of NF-κB (B) and PPARγ (C)
Figure Legend Snippet: Effect of LCE on PPARγ and NF-κB protein levels in mice liver tissue. (A) Measurement of PPARγ and NF-κB protein levels in mice liver were analyzed by western blot. Expression of NF-κB (B) and PPARγ (C)

Techniques Used: Mouse Assay, Western Blot, Expressing

59) Product Images from "The effect of mesoporous bioglass on osteogenesis and adipogenesis of osteoporotic BMSCs"

Article Title: The effect of mesoporous bioglass on osteogenesis and adipogenesis of osteoporotic BMSCs

Journal: Journal of biomedical materials research. Part A

doi: 10.1002/jbm.a.35841

The protein expression of PPARγ and Runx2 in sham and OVX BMSCs cultured in different concentration MBGs dissolution medium (0, 25, 50, 100, and 200 μg/mL) for 14 days (A). Densitometric analysis for the protein expression of Runx2 (B). Densitometric analysis for the protein expression of PPARγ (C).
Figure Legend Snippet: The protein expression of PPARγ and Runx2 in sham and OVX BMSCs cultured in different concentration MBGs dissolution medium (0, 25, 50, 100, and 200 μg/mL) for 14 days (A). Densitometric analysis for the protein expression of Runx2 (B). Densitometric analysis for the protein expression of PPARγ (C).

Techniques Used: Expressing, Cell Culture, Concentration Assay

Real-time PCR analysis of gene Runx2 and PPARγ in sham and OVX BMSCs in different concentration of MBG extract medium (0, 25, 50, 100, and 200 μg/mL) under osteogenic induction for 7 (A, C) of 14 days (B, D). ( n = 3 per group; * p
Figure Legend Snippet: Real-time PCR analysis of gene Runx2 and PPARγ in sham and OVX BMSCs in different concentration of MBG extract medium (0, 25, 50, 100, and 200 μg/mL) under osteogenic induction for 7 (A, C) of 14 days (B, D). ( n = 3 per group; * p

Techniques Used: Real-time Polymerase Chain Reaction, Concentration Assay

60) Product Images from "SIRT1 is regulated by a PPAR?-SIRT1 negative feedback loop associated with senescence"

Article Title: SIRT1 is regulated by a PPAR?-SIRT1 negative feedback loop associated with senescence

Journal: Nucleic Acids Research

doi: 10.1093/nar/gkq609

SIRT1 deacetylase activity influenced appearance of senescence-associated features in 2BS cells. 2BS cells infected with the plasmids pBABE (puro) and pSuper(neo) or pBABE-SIRT1 and pSuper-PPARγshRNA or pBABE-SIRT1 H363Y and pSuper-PPARγshRNA were analyzed for β-gal sataining which is a classical biomarker for senescence. ( A ) Western blot analysis of the SIRT1 and PPARγ protein levels in 2BS cells exposed to the indicated perturbations. Western blotting was performed using specific antibodies against SIRT1 and PPARγ as indicated. The β-actin lane is the loading control. ( B ) β-Gal sataining (blue) in 2BS cells exposed to the indicated preturbations. Stably infected 2BS cells expressing empty vector or both SIRT1 and PPARγshRNA or both SIRT1 H363Y and PPARγshRNA, were passaged until one (cells expressing empty vector) underwent senescence. Young 2BS (22PDs) cells were used as a negative control. Senescent 2BS (61PDs) cells were used as a positive control.
Figure Legend Snippet: SIRT1 deacetylase activity influenced appearance of senescence-associated features in 2BS cells. 2BS cells infected with the plasmids pBABE (puro) and pSuper(neo) or pBABE-SIRT1 and pSuper-PPARγshRNA or pBABE-SIRT1 H363Y and pSuper-PPARγshRNA were analyzed for β-gal sataining which is a classical biomarker for senescence. ( A ) Western blot analysis of the SIRT1 and PPARγ protein levels in 2BS cells exposed to the indicated perturbations. Western blotting was performed using specific antibodies against SIRT1 and PPARγ as indicated. The β-actin lane is the loading control. ( B ) β-Gal sataining (blue) in 2BS cells exposed to the indicated preturbations. Stably infected 2BS cells expressing empty vector or both SIRT1 and PPARγshRNA or both SIRT1 H363Y and PPARγshRNA, were passaged until one (cells expressing empty vector) underwent senescence. Young 2BS (22PDs) cells were used as a negative control. Senescent 2BS (61PDs) cells were used as a positive control.

Techniques Used: Histone Deacetylase Assay, Activity Assay, Infection, Biomarker Assay, Western Blot, Stable Transfection, Expressing, Plasmid Preparation, Negative Control, Positive Control

PPARγ interacts with the endogenous SIRT1 promoter and recruits SIRT1 in HeLa cells. ( A ) Soluble chromatin was prepared from HeLa cells for ChIP analysis of the SIRT1 promoter using antibodies indicated. Precipitated DNA samples were amplified by PCR using a pair of primers that amplify the –592 bp to –163 bp region with GAPDH as an unrelated primers control. ( B ) Luciferase reporter assay of the SIRT1 promoter in SIRT1 transfected HeLa cells. HeLa cells were transfected with expression plasmids vector and SIRT1 as shown, together with SIRT1–Luc. Luciferase activities were measured 48 h after treatment. Values are the mean ± SD of triplicate data points from a representative experiment ( n = 3), which was repeated three times with similar results. ( C ) Real-time quantitative PCR analysis of endogenous SIRT1 mRNA levels in SIRT1-transfected HeLa cells. HeLa cells were transfected with 10 µg of vector or pCDNA3.1-SIRT1. GAPDH transcript was used as a internal control. The error bar represents 1 SD. ( D ) ChIP-upon-ChIP assay showing the colocalization of PPARγ with SIRT1 at the SIRT1 promoter. Soluble chromatin was first immunoprecipitated with rabbit SIRT1 antibody, and the eluted product was re-immunoprecipitated with rabbit PPARγ antibody. ChIP-upon-ChIP assay was also performed with immunoprecipitation in reverse order, that is, PPARγ ChIP followed by SIRT1ChIP. ( E ) PPARγ recruites endogenous SIRT1 to its promoter in PPARγ knockdown cells in a dose-dependent manner. Real-time quantitative PCR verified the occurrence of recruitment. HeLa cells transfected with 10- or 20-nM PPARγ siRNA (triangle) were processed for ChIP uing SIRT1 antibody. The PCR primers amplified the SIRT1 promoter region as indicated in (A). For negative controls, a sample that did not contain antibody (No Ab) was immunoprecipitated, and antibody against β-actin was used as an unrelated antibody control.
Figure Legend Snippet: PPARγ interacts with the endogenous SIRT1 promoter and recruits SIRT1 in HeLa cells. ( A ) Soluble chromatin was prepared from HeLa cells for ChIP analysis of the SIRT1 promoter using antibodies indicated. Precipitated DNA samples were amplified by PCR using a pair of primers that amplify the –592 bp to –163 bp region with GAPDH as an unrelated primers control. ( B ) Luciferase reporter assay of the SIRT1 promoter in SIRT1 transfected HeLa cells. HeLa cells were transfected with expression plasmids vector and SIRT1 as shown, together with SIRT1–Luc. Luciferase activities were measured 48 h after treatment. Values are the mean ± SD of triplicate data points from a representative experiment ( n = 3), which was repeated three times with similar results. ( C ) Real-time quantitative PCR analysis of endogenous SIRT1 mRNA levels in SIRT1-transfected HeLa cells. HeLa cells were transfected with 10 µg of vector or pCDNA3.1-SIRT1. GAPDH transcript was used as a internal control. The error bar represents 1 SD. ( D ) ChIP-upon-ChIP assay showing the colocalization of PPARγ with SIRT1 at the SIRT1 promoter. Soluble chromatin was first immunoprecipitated with rabbit SIRT1 antibody, and the eluted product was re-immunoprecipitated with rabbit PPARγ antibody. ChIP-upon-ChIP assay was also performed with immunoprecipitation in reverse order, that is, PPARγ ChIP followed by SIRT1ChIP. ( E ) PPARγ recruites endogenous SIRT1 to its promoter in PPARγ knockdown cells in a dose-dependent manner. Real-time quantitative PCR verified the occurrence of recruitment. HeLa cells transfected with 10- or 20-nM PPARγ siRNA (triangle) were processed for ChIP uing SIRT1 antibody. The PCR primers amplified the SIRT1 promoter region as indicated in (A). For negative controls, a sample that did not contain antibody (No Ab) was immunoprecipitated, and antibody against β-actin was used as an unrelated antibody control.

Techniques Used: Chromatin Immunoprecipitation, Amplification, Polymerase Chain Reaction, Luciferase, Reporter Assay, Transfection, Expressing, Plasmid Preparation, Real-time Polymerase Chain Reaction, Immunoprecipitation

SIRT1 deacetylates PPARγ in vivo and in vitro . ( A ) Levels of PPARγ acetylation are increased by the SIRT1 inhibitor sirtinol or siRNA Knockdown of SIRT1. Where indicated, cells were treated for 24 h with 100 μM sirtinol or transfected SIRT1-targeting siRNA oligonucleotides for 24 h. PPARγ was immunoprecipitated (IP) from protein lysates using the Flag-epitope tag, and levels of acetylated PPARγ were determined by western blot (WB). ( B ) SIRT1 and p300 form complexes targeting SIRT1 to promoters. Untransfected HeLa cells were analyzed with sequential ChIP assay using SIRT1 and p300 antibodies. The PCR primers amplified the SIRT1 promoter region from –592 to –163 bp with GAPDH as an unrelated primers control, and antibody against β-actin was used as an unrelated antibody control. ( C ) Expression of p300 stimulates PPARγ acetylation. Levels of acetylated PPARγ were assessed in the presence of co-transfected p300 or an acetyltransferase-impaired mutant (p300DN). ( D ) Levels of PPARγ acetylation as a function of increasing amounts of transfected p300. ( E ) Endogenous PPARγ was acetylated in response to trichostatin A (TSA) for 8 h. Endogenous PPARγ was immunoprecipitated and subjected to western blot using antibodies directed against acetylated-lysine residues. The blot was reprobed for the total amount of PPARγ. ( F ) SIRT1 deacetylates PPARγ in vitro in an NAD-dependent manner. HeLa cells were transfected with expression plasmids encoding Flag-PPARγ and p300, and PPARγ was immunoprecipitated from protein lysates using the Flag-epitope tag. SIRT1 from cells overexpressing SIRT1 was purified by immunoprecipitation and added to immunoprecipitated PPARγ for 1 h in the presence or absence of indicated concentrations of either nicotinamide adenine dinucleotide (NAD) or NAM (nicotinamide). Levels of acetylated PPARγ were determined by western blot analysis using an antibody that recognizes acetyl-lysine (Ac-Lys) residues.
Figure Legend Snippet: SIRT1 deacetylates PPARγ in vivo and in vitro . ( A ) Levels of PPARγ acetylation are increased by the SIRT1 inhibitor sirtinol or siRNA Knockdown of SIRT1. Where indicated, cells were treated for 24 h with 100 μM sirtinol or transfected SIRT1-targeting siRNA oligonucleotides for 24 h. PPARγ was immunoprecipitated (IP) from protein lysates using the Flag-epitope tag, and levels of acetylated PPARγ were determined by western blot (WB). ( B ) SIRT1 and p300 form complexes targeting SIRT1 to promoters. Untransfected HeLa cells were analyzed with sequential ChIP assay using SIRT1 and p300 antibodies. The PCR primers amplified the SIRT1 promoter region from –592 to –163 bp with GAPDH as an unrelated primers control, and antibody against β-actin was used as an unrelated antibody control. ( C ) Expression of p300 stimulates PPARγ acetylation. Levels of acetylated PPARγ were assessed in the presence of co-transfected p300 or an acetyltransferase-impaired mutant (p300DN). ( D ) Levels of PPARγ acetylation as a function of increasing amounts of transfected p300. ( E ) Endogenous PPARγ was acetylated in response to trichostatin A (TSA) for 8 h. Endogenous PPARγ was immunoprecipitated and subjected to western blot using antibodies directed against acetylated-lysine residues. The blot was reprobed for the total amount of PPARγ. ( F ) SIRT1 deacetylates PPARγ in vitro in an NAD-dependent manner. HeLa cells were transfected with expression plasmids encoding Flag-PPARγ and p300, and PPARγ was immunoprecipitated from protein lysates using the Flag-epitope tag. SIRT1 from cells overexpressing SIRT1 was purified by immunoprecipitation and added to immunoprecipitated PPARγ for 1 h in the presence or absence of indicated concentrations of either nicotinamide adenine dinucleotide (NAD) or NAM (nicotinamide). Levels of acetylated PPARγ were determined by western blot analysis using an antibody that recognizes acetyl-lysine (Ac-Lys) residues.

Techniques Used: In Vivo, In Vitro, Transfection, Immunoprecipitation, FLAG-tag, Western Blot, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Amplification, Expressing, Mutagenesis, Purification

The catalytic domain of SIRT1 is required for the physical interaction of PPARγ and SIRT1. ( A ) Glutathione S-transferase (GST) pull-down assay shows that with PPARγ interacts directly with SIRT1 in vitro . Three milligrams of either purified GST or GST-PPARγ protein immobilized on glutathione 4B beads was incubated with in vitro translated SIRT1 for 2 h. After extensive washing, the beads were subjected to SDS–PAGE and probed with monoclonal SIRT1 antibody (upper panel). The lower panel shows the result of probing with monoclonal GST antibody. ( B ) HeLa cells were co-transfected with expression plasmids encoding Flag-tagged full-length PPARγ and deletion mutants of SIRT1. Immunoprecipitation and immunoblotting were performed 48 h post-transfection as indicated (+, present; –, absent).
Figure Legend Snippet: The catalytic domain of SIRT1 is required for the physical interaction of PPARγ and SIRT1. ( A ) Glutathione S-transferase (GST) pull-down assay shows that with PPARγ interacts directly with SIRT1 in vitro . Three milligrams of either purified GST or GST-PPARγ protein immobilized on glutathione 4B beads was incubated with in vitro translated SIRT1 for 2 h. After extensive washing, the beads were subjected to SDS–PAGE and probed with monoclonal SIRT1 antibody (upper panel). The lower panel shows the result of probing with monoclonal GST antibody. ( B ) HeLa cells were co-transfected with expression plasmids encoding Flag-tagged full-length PPARγ and deletion mutants of SIRT1. Immunoprecipitation and immunoblotting were performed 48 h post-transfection as indicated (+, present; –, absent).

Techniques Used: Pull Down Assay, In Vitro, Purification, Incubation, SDS Page, Transfection, Expressing, Immunoprecipitation

PPARγ directly regulates SIRT1 transcription. ( A ) Western blot analysis of SIRT1 and PPARγ expression in vector-transfected and PPARγ-transfected or NC-transfected and PPARγsiRNA-transfected HeLa cells with 20 μM troglitazone or DMSO (vehicle) for 48 h. Western blotting was performed using specific antibodies against SIRT1 and PPARγ as indicated. β-Actin or GAPDH was used as a loading control; NC, negative control. ( B ) Real-time PCR analysis of SIRT1 expression in NC-transfected and PPARγsiRNA-transfected HeLa cells with20 μM troglitazone or DMSO (vehicle) for 48 h. Each experiment was performed at least three times. Each bar depicts data from three independent PCR reactions (mean ± SD). ( C ) HeLa cells were transfected with expression plasmids pcDNA3.1 (vector) or pcDNA–PPARγ (PPARγ) together with wild-type SIRT1–Luc or mutant SIRT1–Luc. ( D ) HeLa cells were transfected with NC and PPARγsiRNA together with wild-type SIRT1–Luc. Cells were subsequently treated with 20 μM troglitazone. Luciferase activity was then measured 48 h after treatment. Values are the mean ±.SD of triplicate data points from a representative experiment ( n = 3), which was repeated three times with similar results.
Figure Legend Snippet: PPARγ directly regulates SIRT1 transcription. ( A ) Western blot analysis of SIRT1 and PPARγ expression in vector-transfected and PPARγ-transfected or NC-transfected and PPARγsiRNA-transfected HeLa cells with 20 μM troglitazone or DMSO (vehicle) for 48 h. Western blotting was performed using specific antibodies against SIRT1 and PPARγ as indicated. β-Actin or GAPDH was used as a loading control; NC, negative control. ( B ) Real-time PCR analysis of SIRT1 expression in NC-transfected and PPARγsiRNA-transfected HeLa cells with20 μM troglitazone or DMSO (vehicle) for 48 h. Each experiment was performed at least three times. Each bar depicts data from three independent PCR reactions (mean ± SD). ( C ) HeLa cells were transfected with expression plasmids pcDNA3.1 (vector) or pcDNA–PPARγ (PPARγ) together with wild-type SIRT1–Luc or mutant SIRT1–Luc. ( D ) HeLa cells were transfected with NC and PPARγsiRNA together with wild-type SIRT1–Luc. Cells were subsequently treated with 20 μM troglitazone. Luciferase activity was then measured 48 h after treatment. Values are the mean ±.SD of triplicate data points from a representative experiment ( n = 3), which was repeated three times with similar results.

Techniques Used: Western Blot, Expressing, Plasmid Preparation, Transfection, Negative Control, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Mutagenesis, Luciferase, Activity Assay

Model for the role of PPARγ in the cellular senescence. Reciprocal-type regulation of PPARγ and SIRT1 is proposed for modulation of cellular senescence in response to environmental factors. PPARγ represses the transcription of SIRT1, SIRT1 deacetylates PPARγ posttranscriptionally and PPARγ trans-inhibits SIRT1.
Figure Legend Snippet: Model for the role of PPARγ in the cellular senescence. Reciprocal-type regulation of PPARγ and SIRT1 is proposed for modulation of cellular senescence in response to environmental factors. PPARγ represses the transcription of SIRT1, SIRT1 deacetylates PPARγ posttranscriptionally and PPARγ trans-inhibits SIRT1.

Techniques Used:

Acetylation levels of PPARγ in young and senescence 2BS and WI-38 cells. PPARγ was immunoprecipitated from protein lysates using PPARγ antibody, and acetylation levels of PPARγ were determined by western blot analysis using an antibody that recognizes acetyl-lysine (Ac-lys) residues.
Figure Legend Snippet: Acetylation levels of PPARγ in young and senescence 2BS and WI-38 cells. PPARγ was immunoprecipitated from protein lysates using PPARγ antibody, and acetylation levels of PPARγ were determined by western blot analysis using an antibody that recognizes acetyl-lysine (Ac-lys) residues.

Techniques Used: Immunoprecipitation, Western Blot

61) Product Images from "Maternal n-3 PUFA supplementation promotes fetal brown adipose tissue development through epigenetic modifications in C57BL/6 mice"

Article Title: Maternal n-3 PUFA supplementation promotes fetal brown adipose tissue development through epigenetic modifications in C57BL/6 mice

Journal: Biochimica et biophysica acta. Molecular and cell biology of lipids

doi: 10.1016/j.bbalip.2018.09.008

Maternal n-3 PUFA supplementation promoted brown adipogenesis via histone modifications at the time of weaning. A . Western blot pattern of epigenetic markers of HDAC1, H3K27Ac, H3K27Me3, H3K9Ac, H3K9me2 (left). Image J was used to determine the relative status of histone acetylation and methylation at the H3K27 and H3K9 (right). B. qPCR analysis of epigenetic enzymes of histone methyltransferase Ehmt1, and demethylases of Jmjd3 and Jmjd1 . C . Western blot pattern of adipocyte proteins of PPARγ, FAS, and aP2 (left). The membrane intensity was quantified by Image J (right). D . Enrichment of H3K27Ac at the promoter region of Ucp1 and Pgc1α by ChIP assay in the HIB1B cells that were differentiated with either vehicle (BSA) or EPA (n=4). All data are expressed as mean ± SEM. *p
Figure Legend Snippet: Maternal n-3 PUFA supplementation promoted brown adipogenesis via histone modifications at the time of weaning. A . Western blot pattern of epigenetic markers of HDAC1, H3K27Ac, H3K27Me3, H3K9Ac, H3K9me2 (left). Image J was used to determine the relative status of histone acetylation and methylation at the H3K27 and H3K9 (right). B. qPCR analysis of epigenetic enzymes of histone methyltransferase Ehmt1, and demethylases of Jmjd3 and Jmjd1 . C . Western blot pattern of adipocyte proteins of PPARγ, FAS, and aP2 (left). The membrane intensity was quantified by Image J (right). D . Enrichment of H3K27Ac at the promoter region of Ucp1 and Pgc1α by ChIP assay in the HIB1B cells that were differentiated with either vehicle (BSA) or EPA (n=4). All data are expressed as mean ± SEM. *p

Techniques Used: Western Blot, Methylation, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation

Maternal n-3 PUFA supplementation was associated with metabolic improvement in the later life of offspring. A. Schematic presentation of experimental design. The pups received maternal control diet (Cont)- or FO-diet (FO) were weaned at 3 week-old, and maintained until 11 week-old with no additional dietary modification. B . Average energy expenditure for 4 days measured by metabolic cages (n=5) C . Average RER (VO 2 /VCO 2 ) values for 4 days measured by using metabolic cages (n=5). D . Core body temperature measured by rectal temperature upon exposure to cold temperature (6°C) for 3 hours (n=6). E . Heat release captured by IR camera upon 3-hour cold exposure. F. Representative images with H E staining of iBAT and iWAT after 24-hour exposure to cold temperature (6°C). Red arrows indicate the emergence of brown-like structure within iWAT. G . Western blot pattern of UCP1 in the iBAT and iWAT after 24-hour cold exposure (left). Relative UCP1 levels were quantified by Image J (right). H . Brown signature gene expressions of Ucp1, Pgc1α, and Prdm16 in the IBAT after 24-hour cold exposure (n=6), I . Gene expression levels of Ucp1, Pgc1α, PPARγ and Scerca2b (a responsible gene for calcium- cycling dependent thermogenesis) in the iWAT after 24-hour cold exposure (n=6). All data are expressed as mean ± SEM. *p
Figure Legend Snippet: Maternal n-3 PUFA supplementation was associated with metabolic improvement in the later life of offspring. A. Schematic presentation of experimental design. The pups received maternal control diet (Cont)- or FO-diet (FO) were weaned at 3 week-old, and maintained until 11 week-old with no additional dietary modification. B . Average energy expenditure for 4 days measured by metabolic cages (n=5) C . Average RER (VO 2 /VCO 2 ) values for 4 days measured by using metabolic cages (n=5). D . Core body temperature measured by rectal temperature upon exposure to cold temperature (6°C) for 3 hours (n=6). E . Heat release captured by IR camera upon 3-hour cold exposure. F. Representative images with H E staining of iBAT and iWAT after 24-hour exposure to cold temperature (6°C). Red arrows indicate the emergence of brown-like structure within iWAT. G . Western blot pattern of UCP1 in the iBAT and iWAT after 24-hour cold exposure (left). Relative UCP1 levels were quantified by Image J (right). H . Brown signature gene expressions of Ucp1, Pgc1α, and Prdm16 in the IBAT after 24-hour cold exposure (n=6), I . Gene expression levels of Ucp1, Pgc1α, PPARγ and Scerca2b (a responsible gene for calcium- cycling dependent thermogenesis) in the iWAT after 24-hour cold exposure (n=6). All data are expressed as mean ± SEM. *p

Techniques Used: Modification, Staining, Western Blot, Expressing

62) Product Images from "Pigment Epithelium Derived Factor Peptide Protects Murine Hepatocytes from Carbon Tetrachloride-Induced Injury"

Article Title: Pigment Epithelium Derived Factor Peptide Protects Murine Hepatocytes from Carbon Tetrachloride-Induced Injury

Journal: PLoS ONE

doi: 10.1371/journal.pone.0157647

Effect of STAT3 inhibitor and PEDF receptor siRNA on hepatocyte apoptosis induced by serum deprivation. (A) Inhibitor of STAT3 prevents the induction of PEDF by the 44-mer. Hepatocytes were pretreated with STAT3 inhibitor or ERK inhibitor (PD98059) or PPARγ antagonist (GW9662) for 2 h and then treated with the 44-mer for 24 h. To examine the role of PEDF receptor on antiapoptotic effect of the 44-mer, hepatocytes were transfected with the indicated siRNA. Two days later, the hepatocytes were starved of serum and exposed to the 44-mer for further 24 h. Subsequently, apoptosis was determined by TUNEL assay. Graphs represent means ± SE (n = 4). * P
Figure Legend Snippet: Effect of STAT3 inhibitor and PEDF receptor siRNA on hepatocyte apoptosis induced by serum deprivation. (A) Inhibitor of STAT3 prevents the induction of PEDF by the 44-mer. Hepatocytes were pretreated with STAT3 inhibitor or ERK inhibitor (PD98059) or PPARγ antagonist (GW9662) for 2 h and then treated with the 44-mer for 24 h. To examine the role of PEDF receptor on antiapoptotic effect of the 44-mer, hepatocytes were transfected with the indicated siRNA. Two days later, the hepatocytes were starved of serum and exposed to the 44-mer for further 24 h. Subsequently, apoptosis was determined by TUNEL assay. Graphs represent means ± SE (n = 4). * P

Techniques Used: Transfection, TUNEL Assay

63) Product Images from "Traditional Chinese medicine Qiliqiangxin attenuates phenylephrine-induced cardiac hypertrophy via upregulating PPARγ and PGC-1α"

Article Title: Traditional Chinese medicine Qiliqiangxin attenuates phenylephrine-induced cardiac hypertrophy via upregulating PPARγ and PGC-1α

Journal: Annals of Translational Medicine

doi: 10.21037/atm.2018.04.14

QLQX prevents the NRVMs from cardiac hypertrophy stimulated by PE and upregulates the expression of PPARγ and PGC-1α. (A) QLQX prevented the increased expression of ANP, BNP by qRT-PCR analysis (n=6); (B) immunofluorescent staining showed that increased cardiomyocyte size was reduced by QLQX. Cell membrane was stained for α-actinin (green) and nuclei were counterstained with DAPI (blue). Scale bars, 100 µm (n=3); (C) expression of PPARγ and PGC-1α was determined by western blot in NRVMs treated with PE and/or QLQX (n=6); (D) expression of PPARγ and PGC-1α were determined by western blot in mice treated with PE and/or QLQX (n=6). *, P
Figure Legend Snippet: QLQX prevents the NRVMs from cardiac hypertrophy stimulated by PE and upregulates the expression of PPARγ and PGC-1α. (A) QLQX prevented the increased expression of ANP, BNP by qRT-PCR analysis (n=6); (B) immunofluorescent staining showed that increased cardiomyocyte size was reduced by QLQX. Cell membrane was stained for α-actinin (green) and nuclei were counterstained with DAPI (blue). Scale bars, 100 µm (n=3); (C) expression of PPARγ and PGC-1α was determined by western blot in NRVMs treated with PE and/or QLQX (n=6); (D) expression of PPARγ and PGC-1α were determined by western blot in mice treated with PE and/or QLQX (n=6). *, P

Techniques Used: Expressing, Pyrolysis Gas Chromatography, Aqueous Normal-phase Chromatography, Quantitative RT-PCR, Staining, Western Blot, Mouse Assay

PGC-1α siRNAs or PPARγ inhibitors eliminate the protective effects of QLQX on cardiomyocyte hypertrophy, and PPARγ agonist does not further increase the protective effects of QLQX. (A) PGC-1α siRNAs significantly reduced the PGC-1α expression in NRVMs. n=4; (B) qRT-PCR analysis of ANP and BNP expression in NRVMs treated with PGC-1α siRNAs, PE and/or QLQX (n=6); (C,D). cell size was qualified using immunofluorescent staining for α-actinin (green), nuclei were counterstained with DAPI (blue). Scale bars, 100 µm; n=3; (E,F) expression of ANP, BNP was determined by qRT-PCR analysis after the treatment (n=6); (G,H) using immunofluorescent staining for α-actinin (green) to evaluate the cardiomyocyte size, nuclei were counterstained with DAPI (blue). Scale bars, 100 µm (n=3). *, P
Figure Legend Snippet: PGC-1α siRNAs or PPARγ inhibitors eliminate the protective effects of QLQX on cardiomyocyte hypertrophy, and PPARγ agonist does not further increase the protective effects of QLQX. (A) PGC-1α siRNAs significantly reduced the PGC-1α expression in NRVMs. n=4; (B) qRT-PCR analysis of ANP and BNP expression in NRVMs treated with PGC-1α siRNAs, PE and/or QLQX (n=6); (C,D). cell size was qualified using immunofluorescent staining for α-actinin (green), nuclei were counterstained with DAPI (blue). Scale bars, 100 µm; n=3; (E,F) expression of ANP, BNP was determined by qRT-PCR analysis after the treatment (n=6); (G,H) using immunofluorescent staining for α-actinin (green) to evaluate the cardiomyocyte size, nuclei were counterstained with DAPI (blue). Scale bars, 100 µm (n=3). *, P

Techniques Used: Pyrolysis Gas Chromatography, Expressing, Quantitative RT-PCR, Aqueous Normal-phase Chromatography, Staining

Cardiomyocyte hypertrophy induces downregulation of PPARγ and its coactivator PGC-1α. (A) qRT-PCR for ANP, BNP mRNA levels in cardiomyocytes after 50 µM PE (phenylephrine) treatment (n=6); (B) immunofluorescent staining for α-actinin (green) to qualify the cardiomyocytes size, nuclei were counterstained with DAPI (blue), scale bars, 100 µm (n=4); (C) western blot to detect the expression of PPARγ and PGC-1α in hypertrophic cardiomyocytes (n=6). *, P
Figure Legend Snippet: Cardiomyocyte hypertrophy induces downregulation of PPARγ and its coactivator PGC-1α. (A) qRT-PCR for ANP, BNP mRNA levels in cardiomyocytes after 50 µM PE (phenylephrine) treatment (n=6); (B) immunofluorescent staining for α-actinin (green) to qualify the cardiomyocytes size, nuclei were counterstained with DAPI (blue), scale bars, 100 µm (n=4); (C) western blot to detect the expression of PPARγ and PGC-1α in hypertrophic cardiomyocytes (n=6). *, P

Techniques Used: Pyrolysis Gas Chromatography, Quantitative RT-PCR, Aqueous Normal-phase Chromatography, Staining, Western Blot, Expressing

64) Product Images from "Peroxisome proliferator‐activated receptor gamma (PPARγ) regulates lactase expression and activity in the gut"

Article Title: Peroxisome proliferator‐activated receptor gamma (PPARγ) regulates lactase expression and activity in the gut

Journal: EMBO Molecular Medicine

doi: 10.15252/emmm.201707795

Dose effect of GED , CLA , pioglitazone, and rosiglitazone on LCT expression in Caco‐2 cells Caco‐2 cells were stimulated with various concentrations of each PPARγ agonist for 24 h as indicated. LCT gene expression was determined by qRT–PCR of corresponding reverse‐transcribed mRNA. Results represent the mean ± SEM (two to three independent experiments in triplicate or sixplicate) of the fold change of LCT gene expression. The expression level measured in control cells, arbitrarily defined as one, was used as reference. NS, not significant. Statistical analysis: two‐tailed nonparametric Mann–Whitney U ‐test.
Figure Legend Snippet: Dose effect of GED , CLA , pioglitazone, and rosiglitazone on LCT expression in Caco‐2 cells Caco‐2 cells were stimulated with various concentrations of each PPARγ agonist for 24 h as indicated. LCT gene expression was determined by qRT–PCR of corresponding reverse‐transcribed mRNA. Results represent the mean ± SEM (two to three independent experiments in triplicate or sixplicate) of the fold change of LCT gene expression. The expression level measured in control cells, arbitrarily defined as one, was used as reference. NS, not significant. Statistical analysis: two‐tailed nonparametric Mann–Whitney U ‐test.

Techniques Used: Expressing, Quantitative RT-PCR, Two Tailed Test, MANN-WHITNEY

PPARγ is a transcriptional regulator of the LCT gene Chromatin immunoprecipitation (ChIP) assay. The top diagram depicts the PPRE predicted by in silico analysis. The picture shows PCR amplification of the 8a–8b fragment in ChIP assay from Caco‐2 cells. Graph bars represent quantification of the 8a–8b fragment by qPCR. Results are expressed as fold enrichment with amplification from control cells defined as 1. Luciferase gene reporter assay in Caco‐2 cells transfected with pGL4Luc‐PromLCT, pGL4Luc‐PromLCT MUT, and pGL4Luc‐PromLCT DEL reporter constructs. Cells transfected with empty pGL4Luc were used as control. Results represent the fold change luciferase activity normalized for protein content. LCT gene expression measured by qPCR and LCT activity in PPARγ knockdown Caco‐2 cells (ShPPAR) compared to control cells (ShLuc). LCT activity in ShPPAR cells compared to ShLuc cells stimulated by GED and CLA. The activity levels measured in control cells were arbitrarily defined as one. Effect of GW9662 on GED‐dependent induction of LCT gene expression in Caco‐2 cells. LCT gene expression was determined by qPCR. Results represent the fold change of LCT gene expression. The expression level measured in control cells (w/o GED and GW9662) was used as reference and defined as 1. LCT mRNA expression in the proximal small intestine of PPARγ ΔIEC mice. Results represent the mean ± SD. LCT mRNA expression and activity in Caco‐2 cells stimulated with fenofibrate compared to control cells (DMSO). PPARα mRNA expression in the small intestine of PPARγ ΔIEC mice (left) and in Caco‐2 ShPPARγ/ShLuc cells (right). For mice results, data represent the mean ± SD. Effect of GW6471 on GED‐, CLA‐, Pio‐, and Rosi‐dependent induction of LCT gene expression in Caco‐2 cells. Results represent the fold change of LCT gene expression. The expression level measured in control cells was used as reference and defined as 1. Data information: Data are expressed as mean ± SEM (two to four independent experiments) (except for panels A, E and mouse data shown in G). Statistical analysis: two‐tailed nonparametric Mann–Whitney U ‐test. *** P
Figure Legend Snippet: PPARγ is a transcriptional regulator of the LCT gene Chromatin immunoprecipitation (ChIP) assay. The top diagram depicts the PPRE predicted by in silico analysis. The picture shows PCR amplification of the 8a–8b fragment in ChIP assay from Caco‐2 cells. Graph bars represent quantification of the 8a–8b fragment by qPCR. Results are expressed as fold enrichment with amplification from control cells defined as 1. Luciferase gene reporter assay in Caco‐2 cells transfected with pGL4Luc‐PromLCT, pGL4Luc‐PromLCT MUT, and pGL4Luc‐PromLCT DEL reporter constructs. Cells transfected with empty pGL4Luc were used as control. Results represent the fold change luciferase activity normalized for protein content. LCT gene expression measured by qPCR and LCT activity in PPARγ knockdown Caco‐2 cells (ShPPAR) compared to control cells (ShLuc). LCT activity in ShPPAR cells compared to ShLuc cells stimulated by GED and CLA. The activity levels measured in control cells were arbitrarily defined as one. Effect of GW9662 on GED‐dependent induction of LCT gene expression in Caco‐2 cells. LCT gene expression was determined by qPCR. Results represent the fold change of LCT gene expression. The expression level measured in control cells (w/o GED and GW9662) was used as reference and defined as 1. LCT mRNA expression in the proximal small intestine of PPARγ ΔIEC mice. Results represent the mean ± SD. LCT mRNA expression and activity in Caco‐2 cells stimulated with fenofibrate compared to control cells (DMSO). PPARα mRNA expression in the small intestine of PPARγ ΔIEC mice (left) and in Caco‐2 ShPPARγ/ShLuc cells (right). For mice results, data represent the mean ± SD. Effect of GW6471 on GED‐, CLA‐, Pio‐, and Rosi‐dependent induction of LCT gene expression in Caco‐2 cells. Results represent the fold change of LCT gene expression. The expression level measured in control cells was used as reference and defined as 1. Data information: Data are expressed as mean ± SEM (two to four independent experiments) (except for panels A, E and mouse data shown in G). Statistical analysis: two‐tailed nonparametric Mann–Whitney U ‐test. *** P

Techniques Used: Chromatin Immunoprecipitation, In Silico, Polymerase Chain Reaction, Amplification, Real-time Polymerase Chain Reaction, Luciferase, Reporter Assay, Transfection, Construct, Activity Assay, Expressing, Mouse Assay, Two Tailed Test, MANN-WHITNEY

PPARγ agonists specifically induce LCT expression and activity in Caco‐2 cells Quantitative PCR (qPCR) analysis of LCT gene expression in stimulated Caco‐2 cells. Cells were stimulated for 24 h with each agonist. Results represent the fold change of LCT gene expression normalized to GAPDH level. The expression level measured in control cells was used as reference and defined as 1. Immunofluorescence staining of Caco‐2 cells for LCT protein (green). Cells were stimulated for 24 h with each agonist. Nuclei are stained with DAPI (blue). Non‐relevant IgG was used as control (“IgG control”). Scale bar, 100 μm. Magnification ×20. Western blot analysis of LCT protein expression from stimulated Caco‐2 cells. Densitometric analysis was used to quantify LCT protein. LCT activity in Caco‐2 cells stimulated for 24 h. Results represent the fold change of LCT activity with respect to the activity measured in control cells arbitrarily defined as 1. Data information: (A, D) Data are expressed as mean ± SEM (two to four independent experiments). Statistical analysis: two‐tailed nonparametric Mann–Whitney U ‐test. *** P
Figure Legend Snippet: PPARγ agonists specifically induce LCT expression and activity in Caco‐2 cells Quantitative PCR (qPCR) analysis of LCT gene expression in stimulated Caco‐2 cells. Cells were stimulated for 24 h with each agonist. Results represent the fold change of LCT gene expression normalized to GAPDH level. The expression level measured in control cells was used as reference and defined as 1. Immunofluorescence staining of Caco‐2 cells for LCT protein (green). Cells were stimulated for 24 h with each agonist. Nuclei are stained with DAPI (blue). Non‐relevant IgG was used as control (“IgG control”). Scale bar, 100 μm. Magnification ×20. Western blot analysis of LCT protein expression from stimulated Caco‐2 cells. Densitometric analysis was used to quantify LCT protein. LCT activity in Caco‐2 cells stimulated for 24 h. Results represent the fold change of LCT activity with respect to the activity measured in control cells arbitrarily defined as 1. Data information: (A, D) Data are expressed as mean ± SEM (two to four independent experiments). Statistical analysis: two‐tailed nonparametric Mann–Whitney U ‐test. *** P

Techniques Used: Expressing, Activity Assay, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Western Blot, Two Tailed Test, MANN-WHITNEY

LCT and PPAR γ genes expression correlate in the rat duodenum and jejunum Comparison of the LCT and PPARγ mRNA levels along the gut of “not weaned” and “weaned” rats. Gene expression level was determined by qPCR of corresponding mRNA. Results represent the mean ± SD of the relative expression normalized to GAPDH level (for each group n = 6). * P
Figure Legend Snippet: LCT and PPAR γ genes expression correlate in the rat duodenum and jejunum Comparison of the LCT and PPARγ mRNA levels along the gut of “not weaned” and “weaned” rats. Gene expression level was determined by qPCR of corresponding mRNA. Results represent the mean ± SD of the relative expression normalized to GAPDH level (for each group n = 6). * P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction

65) Product Images from "Long isoforms of NRF1 negatively regulate adipogenesis via suppression of PPARγ expression"

Article Title: Long isoforms of NRF1 negatively regulate adipogenesis via suppression of PPARγ expression

Journal: Redox Biology

doi: 10.1016/j.redox.2019.101414

Overexpression of L-NRF1-741 suppresses the expression of PPAR γ in 3T3-L1 cells. (A) The mRNA expression of Pparγ in control and various Nrf1 -OE cells under normal culture condition. Cont (Control), 313, 453, 572, 583, and 741 refer to the cells overexpressing indicated isoforms of NRF1. * p
Figure Legend Snippet: Overexpression of L-NRF1-741 suppresses the expression of PPAR γ in 3T3-L1 cells. (A) The mRNA expression of Pparγ in control and various Nrf1 -OE cells under normal culture condition. Cont (Control), 313, 453, 572, 583, and 741 refer to the cells overexpressing indicated isoforms of NRF1. * p

Techniques Used: Over Expression, Expressing

L- Nrf1 -KD 3T3-L1 cells exhibits increased mRNA and protein expression of PPARγ under basal condition and during the early stage of adipogenic differentiation. The mRNA expression of adipogenic factors in Scramble and L- Nrf1 -KD cells maintained in normal growth medium (A) and post DMI treatment for indicated time (B). The mRNA levels were measured by RT-qPCR. * p
Figure Legend Snippet: L- Nrf1 -KD 3T3-L1 cells exhibits increased mRNA and protein expression of PPARγ under basal condition and during the early stage of adipogenic differentiation. The mRNA expression of adipogenic factors in Scramble and L- Nrf1 -KD cells maintained in normal growth medium (A) and post DMI treatment for indicated time (B). The mRNA levels were measured by RT-qPCR. * p

Techniques Used: Expressing, Quantitative RT-PCR

66) Product Images from "Prostaglandin reductase-3 negatively modulates adipogenesis through regulation of PPARγ activity [S]"

Article Title: Prostaglandin reductase-3 negatively modulates adipogenesis through regulation of PPARγ activity [S]

Journal: Journal of Lipid Research

doi: 10.1194/jlr.M037556

15-keto-PGE 2 , a PTGR-3 substrate, promotes adipogenesis through activation of PPARγ. (A) Expression of PTGR and PPAR mRNA during adipocyte differentiation. (B) Measurement of 15-keto-PGE 2 level during adipocyte differentiation. Intracellular 15-keto-PGE
Figure Legend Snippet: 15-keto-PGE 2 , a PTGR-3 substrate, promotes adipogenesis through activation of PPARγ. (A) Expression of PTGR and PPAR mRNA during adipocyte differentiation. (B) Measurement of 15-keto-PGE 2 level during adipocyte differentiation. Intracellular 15-keto-PGE

Techniques Used: Activation Assay, Expressing

PTGR-3 decreases proadipogenic effect of 15-keto-PGE 2 on PPARγ activity. Vector-only and PTGR-3 overexpressing preadipocytes were maintained in induction medium with or without 10 μM 15-keto-PGE 2 treatment for 2 days. After 8 days of adipogenic
Figure Legend Snippet: PTGR-3 decreases proadipogenic effect of 15-keto-PGE 2 on PPARγ activity. Vector-only and PTGR-3 overexpressing preadipocytes were maintained in induction medium with or without 10 μM 15-keto-PGE 2 treatment for 2 days. After 8 days of adipogenic

Techniques Used: Activity Assay, Plasmid Preparation

67) Product Images from "Pioglitazone, a PPARγ agonist, attenuates PDGF-induced vascular smooth muscle cell proliferation through AMPK-dependent and AMPK-independent inhibition of mTOR/p70S6K and ERK signaling"

Article Title: Pioglitazone, a PPARγ agonist, attenuates PDGF-induced vascular smooth muscle cell proliferation through AMPK-dependent and AMPK-independent inhibition of mTOR/p70S6K and ERK signaling

Journal: Biochemical pharmacology

doi: 10.1016/j.bcp.2015.11.026

Effects of PPARγ downregulation or PPARγ inhibition on PIO-induced changes in the phosphorylation of AMPK, S6, and ERK1/2, and cyclin D1 expression in VSMCs. (A) Serum-deprived VSMCs were treated with PIO (30 μM, 48 hr) or vehicle
Figure Legend Snippet: Effects of PPARγ downregulation or PPARγ inhibition on PIO-induced changes in the phosphorylation of AMPK, S6, and ERK1/2, and cyclin D1 expression in VSMCs. (A) Serum-deprived VSMCs were treated with PIO (30 μM, 48 hr) or vehicle

Techniques Used: Inhibition, Expressing

68) Product Images from "Evasion of immunosurveillance by genomic alterations of PPARγ/RXRα in bladder cancer"

Article Title: Evasion of immunosurveillance by genomic alterations of PPARγ/RXRα in bladder cancer

Journal: Nature Communications

doi: 10.1038/s41467-017-00147-w

S427F mutation in RXRα stabilizes heterodimerization with PPARγ and promotes the agonistic conformation. a Sizing profile of RXRα S427F mutant ( green ), PPARγ ( purple ), and the heterodimer ( magenta ). Both RXRα S427F and PPARγ run as monomers. When mixed together in 1:1 stoichiometry, the elution profile shifts demonstrating formation of the heterodimer in the absence of ligand. b SPR demonstrating enhanced interaction between RXRα S427F mutant and PPARγ. RXRα was immobilized to the CM5 chip by amine coupling and PPARγ was injected in dose response from 3 μM to 24 nM with 60 s association phase and 120 s disassociation. c Overall crystal structure of the heterodimer complex of RXRα S427F mutant ( green ) and PPARγ ( blue ) with the co-activator peptide Src1 ( red ). The agonists 9-cis-retinoic acid and rosiglitazone are rendered as spheres. The AF-2 helix (Helix H12) of PPARγ has been highlighted in magenta . RXRα S427 and PPARγ Y477 are rendered as sticks and located in the dimer interface. d Zoom in of the heterodimer interface shows the S427F mutation of RXRα ( green ) introduces a π-stacking interaction with Y477 of PPARg ( blue ) at the C-terminus ( magenta ). The 2Fo–Fc electron density map is shown in gray and contoured at 1.2 s
Figure Legend Snippet: S427F mutation in RXRα stabilizes heterodimerization with PPARγ and promotes the agonistic conformation. a Sizing profile of RXRα S427F mutant ( green ), PPARγ ( purple ), and the heterodimer ( magenta ). Both RXRα S427F and PPARγ run as monomers. When mixed together in 1:1 stoichiometry, the elution profile shifts demonstrating formation of the heterodimer in the absence of ligand. b SPR demonstrating enhanced interaction between RXRα S427F mutant and PPARγ. RXRα was immobilized to the CM5 chip by amine coupling and PPARγ was injected in dose response from 3 μM to 24 nM with 60 s association phase and 120 s disassociation. c Overall crystal structure of the heterodimer complex of RXRα S427F mutant ( green ) and PPARγ ( blue ) with the co-activator peptide Src1 ( red ). The agonists 9-cis-retinoic acid and rosiglitazone are rendered as spheres. The AF-2 helix (Helix H12) of PPARγ has been highlighted in magenta . RXRα S427 and PPARγ Y477 are rendered as sticks and located in the dimer interface. d Zoom in of the heterodimer interface shows the S427F mutation of RXRα ( green ) introduces a π-stacking interaction with Y477 of PPARg ( blue ) at the C-terminus ( magenta ). The 2Fo–Fc electron density map is shown in gray and contoured at 1.2 s

Techniques Used: Mutagenesis, SPR Assay, Chromatin Immunoprecipitation, Injection

Tumor-intrinsic activation of PPARγ/RXRα is negatively correlated with immune infiltration. a Pathway enrichment analysis of genes differentially expressed in RXRA-S427Y, RXRA-S427F and PPARG overexpressing T24 lines relative to respective controls. Top suppressed pathways are shown. The analysis was based on three biological replicates. b Dot plot showing expression correlation of all genes with the curated immune signature (refer to “Methods”) vs. correlation with PPARG in bladder tumors ( n = 385) from TCGA. c Heatmap presenting associations between RXRA mutations and PPARG expression with T-cell markers ( top, green label ), immune checkpoint molecules ( middle, yellow label ), and pro-inflammatory factors ( bottom, lavender label ) in TCGA MIBC ( n = 385). d IHC staining of PPARγ and CD8 in two representative human bladder tumor samples from a clinical cohort ( n = 23, Eisai cohort). Scale bars: 100 μm. e Summary of the IHC results of Eisai cohort shown in d . Distribution of CD8+ T-cell infiltration in bladder tumors expressing high (scores 2–4) or low (score 1) levels of PPARγ protein. f Whisker plot representing IHC staining of infiltrating CD8+ T cells and PPARγ protein expression of MIBC samples from the bladder cancer meta-dataset ( n = 118). No expression, score = 1; High expression, score = 4. The bold lines: median; the boxes: interquartile range ( IQR ); the upper whiskers: min(max(x), Q_3 + 1.5 * IQR); the lower whiskers: max(min(x), Q_1−1.5 * IQR). Statistical analysis was performed using Kruskal–Wallis test
Figure Legend Snippet: Tumor-intrinsic activation of PPARγ/RXRα is negatively correlated with immune infiltration. a Pathway enrichment analysis of genes differentially expressed in RXRA-S427Y, RXRA-S427F and PPARG overexpressing T24 lines relative to respective controls. Top suppressed pathways are shown. The analysis was based on three biological replicates. b Dot plot showing expression correlation of all genes with the curated immune signature (refer to “Methods”) vs. correlation with PPARG in bladder tumors ( n = 385) from TCGA. c Heatmap presenting associations between RXRA mutations and PPARG expression with T-cell markers ( top, green label ), immune checkpoint molecules ( middle, yellow label ), and pro-inflammatory factors ( bottom, lavender label ) in TCGA MIBC ( n = 385). d IHC staining of PPARγ and CD8 in two representative human bladder tumor samples from a clinical cohort ( n = 23, Eisai cohort). Scale bars: 100 μm. e Summary of the IHC results of Eisai cohort shown in d . Distribution of CD8+ T-cell infiltration in bladder tumors expressing high (scores 2–4) or low (score 1) levels of PPARγ protein. f Whisker plot representing IHC staining of infiltrating CD8+ T cells and PPARγ protein expression of MIBC samples from the bladder cancer meta-dataset ( n = 118). No expression, score = 1; High expression, score = 4. The bold lines: median; the boxes: interquartile range ( IQR ); the upper whiskers: min(max(x), Q_3 + 1.5 * IQR); the lower whiskers: max(min(x), Q_1−1.5 * IQR). Statistical analysis was performed using Kruskal–Wallis test

Techniques Used: Activation Assay, Expressing, Immunohistochemistry, Staining, Whisker Assay

RXRα S427F/Y functionally promotes ligand-independent PPARγ signaling in human bladder cancer lines. a Heat map representing pathways activated/suppressed in RXRα S427Y , RXRα S427F and PPARγ overexpressing lines relative to their respective controls. Orange represents pathway activation and blue represents pathway suppression. The analysis was based on three biological replicates. b Upper , western blot of RXRα confirming overexpression of RXRα WT (WT), RXRα S427F (S427F) and RXRα S427Y (S427Y) in T24 cells relative to control (Vec). Lower , RT-qPCR analysis of ANGPTL4 and PLIN2 in various engineered lines. c Upper , western blot confirming overexpression of PPARγ in T24 line relative to control (Vec). Lower , RT-qPCR analysis of ANGPTL4 , PLIN2 , ACOX1 and PDK4 in engineered lines. d Upper , western blot of RXRα and PPARγ in SV-HUC line engineered to inducibly overexpress RXRα S427F and knockdown PPARγ by multiple shRNAs (sh#4, 5 and 9) upon doxycycline ( DOX ) treatment. Lower , RT-qPCR analysis of PLIN2 , ACOX1 and PSCA in various SV-HUC-1 engineered lines. +/− represents presence or absence of DOX treatment respectively. e RT-qPCR analysis of ANGPTL4 and PLIN2 in HT-1197 (carrying endogenous RXRA S427F ), 5637 and UM-UC9 (PPARG amplified) lines treated with DMSO or T0070907 for 24 h. All RT-qPCR data is normalized to GAPDH and presented as mean fold change vs. control ± SEM of at least three biological replicates
Figure Legend Snippet: RXRα S427F/Y functionally promotes ligand-independent PPARγ signaling in human bladder cancer lines. a Heat map representing pathways activated/suppressed in RXRα S427Y , RXRα S427F and PPARγ overexpressing lines relative to their respective controls. Orange represents pathway activation and blue represents pathway suppression. The analysis was based on three biological replicates. b Upper , western blot of RXRα confirming overexpression of RXRα WT (WT), RXRα S427F (S427F) and RXRα S427Y (S427Y) in T24 cells relative to control (Vec). Lower , RT-qPCR analysis of ANGPTL4 and PLIN2 in various engineered lines. c Upper , western blot confirming overexpression of PPARγ in T24 line relative to control (Vec). Lower , RT-qPCR analysis of ANGPTL4 , PLIN2 , ACOX1 and PDK4 in engineered lines. d Upper , western blot of RXRα and PPARγ in SV-HUC line engineered to inducibly overexpress RXRα S427F and knockdown PPARγ by multiple shRNAs (sh#4, 5 and 9) upon doxycycline ( DOX ) treatment. Lower , RT-qPCR analysis of PLIN2 , ACOX1 and PSCA in various SV-HUC-1 engineered lines. +/− represents presence or absence of DOX treatment respectively. e RT-qPCR analysis of ANGPTL4 and PLIN2 in HT-1197 (carrying endogenous RXRA S427F ), 5637 and UM-UC9 (PPARG amplified) lines treated with DMSO or T0070907 for 24 h. All RT-qPCR data is normalized to GAPDH and presented as mean fold change vs. control ± SEM of at least three biological replicates

Techniques Used: Activation Assay, Western Blot, Over Expression, Quantitative RT-PCR, Amplification

PPARγ/RXRα S427F confers partial resistance to immunotherapies. a RT-qPCR analysis of chemokines/cytokines in T24 lines engineered to overexpress RXRA-WT, RXRA-S427F, RXRA-S427Y ( upper ), and PPARG ( lower ). Controls are RXRA-WT for RXRA mutant lines and vector control (Vec) for PPARG overexpressing line. Expression normalized to GAPDH and data presented as mean fold change vs. control ± SEM of three biological replicates. b Chemokine array analysis of conditioned media collected from T24 lines engineered to overexpress PPARG (PPARγ) vs. control (Vec). Dotted boxes represent controls. Significant changes in secretion are outlined. One representative of three independent experiments is shown. c FACS based quantitation of infiltrating CD3 + CD8 + double positive T cells into subcutaneously implanted MBT2 tumors overexpressing RXRA-WT ( n = 6) or RXRA-S427F ( n = 6). Data presented as percent of total tumor-derived cells following dissociation. d Left , individual MBT2-RXRα WT tumor volumes in response to PBS ( red , n = 12) or anti-CTLA4 ( blue , n = 12). P = 0.0189 at day 7. Right , individual MBT2-RXRα S427F tumor volumes in response to PBS ( red , n = 12) or anti-CTLA4 ( blue , n = 12). P > 0.05 at day 7. One-way ANOVA followed by Tukey’s post-hoc test performed. e Heatmap presenting pathway level analysis (activation, red ; suppression, blue ) of differentially expressed genes in PPARγ knockdown lines (PPARγ-sh#4 and -sh#9 engineered in SV-HUC-1 line expressing RXRA-S427Y) relative to vector control. The analysis was based on three biological replicates. f Left , Knockdown of PPARγ or RXRα by shRNAs in HT-1197 cells. GAPDH was used as the control. Right , RT-qPCR analysis of inflammatory genes CCL2 and CXCL10 following inducible knockdown of PPARγ and RXRα in HT-1197 cells. Data normalized to GAPDH and presented as mean fold change (Dox treated vs. untreated) ± SEM of three biological replicates. g RT-qPCR analysis of IL8 and CCL2 following treatment with PPARγ agonist rosiglitazone (Rosi) or PPARγ antagonist T0070907 in 5637 cells. Data normalized to GAPDH and presented as mean fold change ± SEM of three biological replicates. h Schematic representation of the role of tumor-intrinsic PPARγ/RXRα S427F/Y in transcriptional regulation and immunosurveillance. CoA, co-activator complex; CoR, co-repressor complex; ITF, inflammation-related transcription factors
Figure Legend Snippet: PPARγ/RXRα S427F confers partial resistance to immunotherapies. a RT-qPCR analysis of chemokines/cytokines in T24 lines engineered to overexpress RXRA-WT, RXRA-S427F, RXRA-S427Y ( upper ), and PPARG ( lower ). Controls are RXRA-WT for RXRA mutant lines and vector control (Vec) for PPARG overexpressing line. Expression normalized to GAPDH and data presented as mean fold change vs. control ± SEM of three biological replicates. b Chemokine array analysis of conditioned media collected from T24 lines engineered to overexpress PPARG (PPARγ) vs. control (Vec). Dotted boxes represent controls. Significant changes in secretion are outlined. One representative of three independent experiments is shown. c FACS based quantitation of infiltrating CD3 + CD8 + double positive T cells into subcutaneously implanted MBT2 tumors overexpressing RXRA-WT ( n = 6) or RXRA-S427F ( n = 6). Data presented as percent of total tumor-derived cells following dissociation. d Left , individual MBT2-RXRα WT tumor volumes in response to PBS ( red , n = 12) or anti-CTLA4 ( blue , n = 12). P = 0.0189 at day 7. Right , individual MBT2-RXRα S427F tumor volumes in response to PBS ( red , n = 12) or anti-CTLA4 ( blue , n = 12). P > 0.05 at day 7. One-way ANOVA followed by Tukey’s post-hoc test performed. e Heatmap presenting pathway level analysis (activation, red ; suppression, blue ) of differentially expressed genes in PPARγ knockdown lines (PPARγ-sh#4 and -sh#9 engineered in SV-HUC-1 line expressing RXRA-S427Y) relative to vector control. The analysis was based on three biological replicates. f Left , Knockdown of PPARγ or RXRα by shRNAs in HT-1197 cells. GAPDH was used as the control. Right , RT-qPCR analysis of inflammatory genes CCL2 and CXCL10 following inducible knockdown of PPARγ and RXRα in HT-1197 cells. Data normalized to GAPDH and presented as mean fold change (Dox treated vs. untreated) ± SEM of three biological replicates. g RT-qPCR analysis of IL8 and CCL2 following treatment with PPARγ agonist rosiglitazone (Rosi) or PPARγ antagonist T0070907 in 5637 cells. Data normalized to GAPDH and presented as mean fold change ± SEM of three biological replicates. h Schematic representation of the role of tumor-intrinsic PPARγ/RXRα S427F/Y in transcriptional regulation and immunosurveillance. CoA, co-activator complex; CoR, co-repressor complex; ITF, inflammation-related transcription factors

Techniques Used: Quantitative RT-PCR, Mutagenesis, Plasmid Preparation, Expressing, FACS, Quantitation Assay, Derivative Assay, Activation Assay

69) Product Images from "Inhibitory effects of Orostachys malacophyllus var. iwarenge extracts on reactive oxygen species production and lipid accumulation during 3T3-L1 adipocyte differentiation"

Article Title: Inhibitory effects of Orostachys malacophyllus var. iwarenge extracts on reactive oxygen species production and lipid accumulation during 3T3-L1 adipocyte differentiation

Journal: Food Science and Biotechnology

doi: 10.1007/s10068-018-0426-x

Effects of O. malacophyllus extracts on mRNA and protein expression of adipogenic transcription factors. 3T3-L1 cells were treated with O. malacophyllus extract (10, 50, and 100 μg/mL) during adipogenesis. The control was adipocytes differentiated for 8 days with MDI, a hormone mixture. NAC was the positive control. Total RNA was extracted on day 8, and ( A ) PPARγ, ( B ) C/EBPα and ( C ) aP2 mRNA were measured using quantitative real-time PCR. Total protein levels were measured using Western blotting with ( D ) PPARγ, ( E ) C/EBPα and ( F ) aP2 antibodies. mRNA and protein levels were quantified and normalized to that of β-actin. The band intensities were measured with software Imag. Each value represents the mean ± standard deviation of triplicate experiments. Values with different letters indicate statistically significant differences among groups at p
Figure Legend Snippet: Effects of O. malacophyllus extracts on mRNA and protein expression of adipogenic transcription factors. 3T3-L1 cells were treated with O. malacophyllus extract (10, 50, and 100 μg/mL) during adipogenesis. The control was adipocytes differentiated for 8 days with MDI, a hormone mixture. NAC was the positive control. Total RNA was extracted on day 8, and ( A ) PPARγ, ( B ) C/EBPα and ( C ) aP2 mRNA were measured using quantitative real-time PCR. Total protein levels were measured using Western blotting with ( D ) PPARγ, ( E ) C/EBPα and ( F ) aP2 antibodies. mRNA and protein levels were quantified and normalized to that of β-actin. The band intensities were measured with software Imag. Each value represents the mean ± standard deviation of triplicate experiments. Values with different letters indicate statistically significant differences among groups at p

Techniques Used: Expressing, Positive Control, Real-time Polymerase Chain Reaction, Western Blot, Software, Standard Deviation

70) Product Images from "PPARγ in dendritic cells and T cells drives pathogenic type-2 effector responses in lung inflammation"

Article Title: PPARγ in dendritic cells and T cells drives pathogenic type-2 effector responses in lung inflammation

Journal: The Journal of Experimental Medicine

doi: 10.1084/jem.20162069

PPARγ in DCs intrinsically controls Th2 polarization. Lung DCs from Pparg fl/fl and Cd11c -Cre Pparg fl/fl mice, which had been reconstituted with WT AMs were sorted and cultured in vitro for 4 d with OTII cells and 10 nM OVA 323-339 peptide. Frequency of IL-4 + , IFNγ + and IL-17A + CD4 + T cells (A) and the proliferation of efluor-670 labeled OTII cells (B; n = 3/condition) are shown. (C and D) Pparg fl/fl and Cd11c -Cre Pparg fl/fl mice, which had been reconstituted with WT AMs were injected with 100 µg HDM. 24 h after infection, the lung conventional DC subsets of each genotype were sorted and transferred intratracheally to naive WT recipients. After 10 d, those animals were challenged for a consecutive 5 d with 10 µg HDM and were subsequently analyzed on day 17. Total number of eosinophils and neutrophils (C) and the total numbers of cytokine producing CD4 + T cells in the BAL (D; n = 5/group). Data are means ± SEM and the sample size ( n ) and are representative of two independent experiments. ANOVA (one-way) was used. *, P
Figure Legend Snippet: PPARγ in DCs intrinsically controls Th2 polarization. Lung DCs from Pparg fl/fl and Cd11c -Cre Pparg fl/fl mice, which had been reconstituted with WT AMs were sorted and cultured in vitro for 4 d with OTII cells and 10 nM OVA 323-339 peptide. Frequency of IL-4 + , IFNγ + and IL-17A + CD4 + T cells (A) and the proliferation of efluor-670 labeled OTII cells (B; n = 3/condition) are shown. (C and D) Pparg fl/fl and Cd11c -Cre Pparg fl/fl mice, which had been reconstituted with WT AMs were injected with 100 µg HDM. 24 h after infection, the lung conventional DC subsets of each genotype were sorted and transferred intratracheally to naive WT recipients. After 10 d, those animals were challenged for a consecutive 5 d with 10 µg HDM and were subsequently analyzed on day 17. Total number of eosinophils and neutrophils (C) and the total numbers of cytokine producing CD4 + T cells in the BAL (D; n = 5/group). Data are means ± SEM and the sample size ( n ) and are representative of two independent experiments. ANOVA (one-way) was used. *, P

Techniques Used: Mouse Assay, Affinity Magnetic Separation, Cell Culture, In Vitro, Labeling, Injection, Infection

IL-4 receptor and IL-33 receptor signaling controls PPARγ expression and DC-mediated Th2 polarization. (A) PPARγ expression in lung DCs from naive WT and Il1rl1 −/− mice was measured using flow cytometry. Shown is the MFI of PPARγ normalized to the PPARγ-deficient control of the indicated DC subsets ( n = 3/group). (B) Lung DCs from WT and Il1rl1 −/− animals were isolated and co-cultured in vitro for 4 d with naive, splenic, Smarta-1, transgenic CD4 + T cells and 1 nM gp61 peptide. Frequency of IL-4 + and IFN-γ + CD4 + T cells ( n = 10/condition). (C) Surface expression of ST2 on WT lung and splenic DCs was evaluated using flow cytometry. Shown are representative FACS plots including fluorescence minus one (FMO) controls ( n = 3/group). (D and E) WT animals received PBS or 100 µg HDM intratracheally and lungs were analyzed for cytokine-producing cells 18 h after infection. Shown is the frequency of cytokine-producing ILC2s (D) and basophils (E) of indicated cytokines ( n = 4/group). (F) PPARγ expression in lung DCs from naive WT and Il4ra −/− mice was measured using flow cytometry. Shown is the MFI of PPARγ normalized to the PPARγ-deficient control of the indicated DC subsets ( n = 3/group). (G) Lung DCs from WT and Il4ra −/− animals were isolated and co-cultured in vitro for 4 d with naive, splenic, Smarta-1, transgenic CD4 + T cells and 1 nM gp61 peptide. Shown are the frequencies of IL-4 + , IL-5 + , IL-13 + , and IFN-γ + CD4 + T cells ( n = 8/condition) Data are means ± SEM and the sample size ( n ). ANOVA (one way) was used. *, P
Figure Legend Snippet: IL-4 receptor and IL-33 receptor signaling controls PPARγ expression and DC-mediated Th2 polarization. (A) PPARγ expression in lung DCs from naive WT and Il1rl1 −/− mice was measured using flow cytometry. Shown is the MFI of PPARγ normalized to the PPARγ-deficient control of the indicated DC subsets ( n = 3/group). (B) Lung DCs from WT and Il1rl1 −/− animals were isolated and co-cultured in vitro for 4 d with naive, splenic, Smarta-1, transgenic CD4 + T cells and 1 nM gp61 peptide. Frequency of IL-4 + and IFN-γ + CD4 + T cells ( n = 10/condition). (C) Surface expression of ST2 on WT lung and splenic DCs was evaluated using flow cytometry. Shown are representative FACS plots including fluorescence minus one (FMO) controls ( n = 3/group). (D and E) WT animals received PBS or 100 µg HDM intratracheally and lungs were analyzed for cytokine-producing cells 18 h after infection. Shown is the frequency of cytokine-producing ILC2s (D) and basophils (E) of indicated cytokines ( n = 4/group). (F) PPARγ expression in lung DCs from naive WT and Il4ra −/− mice was measured using flow cytometry. Shown is the MFI of PPARγ normalized to the PPARγ-deficient control of the indicated DC subsets ( n = 3/group). (G) Lung DCs from WT and Il4ra −/− animals were isolated and co-cultured in vitro for 4 d with naive, splenic, Smarta-1, transgenic CD4 + T cells and 1 nM gp61 peptide. Shown are the frequencies of IL-4 + , IL-5 + , IL-13 + , and IFN-γ + CD4 + T cells ( n = 8/condition) Data are means ± SEM and the sample size ( n ). ANOVA (one way) was used. *, P

Techniques Used: Expressing, Mouse Assay, Flow Cytometry, Cytometry, Isolation, Cell Culture, In Vitro, Transgenic Assay, FACS, Fluorescence, Infection

PPARγ is highly expressed in mouse T cells and controls Th2 polarization in vitro . Naive splenic CD4 + T cells were sorted from Pparg fl/fl and Cd4 -Cre Pparg fl/fl mice and were co-cultured for 4 d with splenic DCs and soluble αCD3 mAbs (2 µg/ml). Shown are the Pparg mRNA of Pparg fl/fl cells (WT; A) and the intracellular protein expression of PPARγ in Pparg fl/fl cells (B), with the corresponding Cd4 -Cre Pparg fl/fl controls and PPARγ expression in AMs as a comparison (C). (D) Cytokine production profile after 4 h restimulation with PMA/ionomycin ( n = 4/condition). (E and F) naive, splenic, Smarta-1, transgenic CD4 + T cells were co-cultured with splenic DCs and 10 nM gp61 peptide in polarizing conditions in the presence of PPARγ antagonist GW9662 at the indicated concentrations or in vehicle (DMSO) as a control. Shown are the frequencies of ST2 + and cytokine-producing cells of indicated cytokines after restimulation with PMA/ionomycin. (E) GW9662 was added from the beginning and cells were analyzed after 4 d of culture ( n = 4/condition). (F) Cells were cultured for 4 d before addition of GW9662 and analyzed 24 h later ( n = 4/condition). The data are representative of three independent experiments. Data are means ± SEM and the sample size ( n ). ANOVA (one-way) was used. *, P
Figure Legend Snippet: PPARγ is highly expressed in mouse T cells and controls Th2 polarization in vitro . Naive splenic CD4 + T cells were sorted from Pparg fl/fl and Cd4 -Cre Pparg fl/fl mice and were co-cultured for 4 d with splenic DCs and soluble αCD3 mAbs (2 µg/ml). Shown are the Pparg mRNA of Pparg fl/fl cells (WT; A) and the intracellular protein expression of PPARγ in Pparg fl/fl cells (B), with the corresponding Cd4 -Cre Pparg fl/fl controls and PPARγ expression in AMs as a comparison (C). (D) Cytokine production profile after 4 h restimulation with PMA/ionomycin ( n = 4/condition). (E and F) naive, splenic, Smarta-1, transgenic CD4 + T cells were co-cultured with splenic DCs and 10 nM gp61 peptide in polarizing conditions in the presence of PPARγ antagonist GW9662 at the indicated concentrations or in vehicle (DMSO) as a control. Shown are the frequencies of ST2 + and cytokine-producing cells of indicated cytokines after restimulation with PMA/ionomycin. (E) GW9662 was added from the beginning and cells were analyzed after 4 d of culture ( n = 4/condition). (F) Cells were cultured for 4 d before addition of GW9662 and analyzed 24 h later ( n = 4/condition). The data are representative of three independent experiments. Data are means ± SEM and the sample size ( n ). ANOVA (one-way) was used. *, P

Techniques Used: In Vitro, Mouse Assay, Cell Culture, Expressing, Affinity Magnetic Separation, Transgenic Assay

PPARγ is largely dispensable for lung DC activation and antigen uptake but mediates antigen transport by CD11b + DCs to the dLN. WT and AM-reconstituted Cd11c -Cre Pparg fl/fl were injected intratracheally with 100 µg OVA-AF488 and 100 µg HDM and sacrificed 24 h later for analysis of lung and dLNs by flow cytometry. DC subsets were identified as CD45 + Siglec-F − CD11c + MHCII high for the lung and CD45 + autofluorescent − CD11c + MHCII high for the lung and dLNs, respectively. Frequency of OVA-AF488 + among each DC subset in the lung dLN (A) and the total number of OVA-AF488 + DCs (B). Frequency of OVA-AF488 + among each DC subset in the lung (C) and the total the total number of OVA-AF488 + DCs (D). MFIs of indicated DC activation markers for lung dLN (E–G) and lung (H–J) DC subsets as a summary of FACS data. (K and L) MFI of ST2-expressing cells among DCs in the lung (K) and the lung dLN (L; n = 4–6/group). The data are representative of two experiments and are means ± SEM. The Student’s t test (unpaired) was used. *, P
Figure Legend Snippet: PPARγ is largely dispensable for lung DC activation and antigen uptake but mediates antigen transport by CD11b + DCs to the dLN. WT and AM-reconstituted Cd11c -Cre Pparg fl/fl were injected intratracheally with 100 µg OVA-AF488 and 100 µg HDM and sacrificed 24 h later for analysis of lung and dLNs by flow cytometry. DC subsets were identified as CD45 + Siglec-F − CD11c + MHCII high for the lung and CD45 + autofluorescent − CD11c + MHCII high for the lung and dLNs, respectively. Frequency of OVA-AF488 + among each DC subset in the lung dLN (A) and the total number of OVA-AF488 + DCs (B). Frequency of OVA-AF488 + among each DC subset in the lung (C) and the total the total number of OVA-AF488 + DCs (D). MFIs of indicated DC activation markers for lung dLN (E–G) and lung (H–J) DC subsets as a summary of FACS data. (K and L) MFI of ST2-expressing cells among DCs in the lung (K) and the lung dLN (L; n = 4–6/group). The data are representative of two experiments and are means ± SEM. The Student’s t test (unpaired) was used. *, P

Techniques Used: Activation Assay, Injection, Flow Cytometry, Cytometry, FACS, Expressing

PPARγ in CD11c + cells intrinsically mediates pulmonary allergic inflammation. (A–D) Pparg fl/fl and Cd11c -Cre Pparg fl/fl mice were sensitized and challenged intratracheally (i.t.), as indicated in the scheme with HDM extract. Total cell numbers in the BAL of eosinophils, neutrophils and AMs (B), CD4 + and CD8 + T cells (C), and the total amount of IgE in the BAL (D; n = 5/group). (E–G) WT and Cd11c -Cre Pparg fl/fl mice sensitized with OVA/alum i.p. and challenged i.t. A scheme of the immunization protocol (E); total cell numbers in the BAL of eosinophils, neutrophils, and AMs (F); and the total number of CD4 + T cells in the BAL (G; n = 5/group). (H–K) AMs were depleted with clodronate before HDM sensitization and challenge, as depicted in the scheme (H). The number of lung eosinophils and neutrophils (I) and CD4 + and CD8 + T cells (J). (K) Depicted is the frequency of cytokine-producing cells of CD4 + T cells after restimulation with PMA/ionomycin ( n = 4/group). The data are representative of two experiments each and are means ± SEM, with the sample size ( n ). The Student’s t test (unpaired) was used. *, P
Figure Legend Snippet: PPARγ in CD11c + cells intrinsically mediates pulmonary allergic inflammation. (A–D) Pparg fl/fl and Cd11c -Cre Pparg fl/fl mice were sensitized and challenged intratracheally (i.t.), as indicated in the scheme with HDM extract. Total cell numbers in the BAL of eosinophils, neutrophils and AMs (B), CD4 + and CD8 + T cells (C), and the total amount of IgE in the BAL (D; n = 5/group). (E–G) WT and Cd11c -Cre Pparg fl/fl mice sensitized with OVA/alum i.p. and challenged i.t. A scheme of the immunization protocol (E); total cell numbers in the BAL of eosinophils, neutrophils, and AMs (F); and the total number of CD4 + T cells in the BAL (G; n = 5/group). (H–K) AMs were depleted with clodronate before HDM sensitization and challenge, as depicted in the scheme (H). The number of lung eosinophils and neutrophils (I) and CD4 + and CD8 + T cells (J). (K) Depicted is the frequency of cytokine-producing cells of CD4 + T cells after restimulation with PMA/ionomycin ( n = 4/group). The data are representative of two experiments each and are means ± SEM, with the sample size ( n ). The Student’s t test (unpaired) was used. *, P

Techniques Used: Mouse Assay, Affinity Magnetic Separation

PPARγ suppresses a Th17/Th22 transcriptional program and is essential for IL-33–induced CD4 + T cell effector cytokine production. Naive splenic Smarta-1 transgenic CD4 + T cells were co-cultured with splenic DCs and 10 nM gp33 peptide in Th2 polarizing conditions in the presence of PPARγ antagonist GW9662 or vehicle (DMSO) as a control. After 4 d of co-culture, RNA sequencing of CD4 + T cells was performed ( n = 2). Heat map of the 100 most-differentially regulated genes (A) and expression levels of selected Th2 cytokines (B). (C) Naive splenic Smarta-1 transgenic CD4 + T cells were co-cultured with splenic DCs and 10 nM gp61 peptide in Th2 or Th22 polarizing conditions in the presence of PPARγ antagonist GW9662 or vehicle (DMSO) as a control. (C and D) Shown is the production of IL-17F (C) and IL-22 (D) after 4-h restimulation with PMA/ionomycin ( n = 4/condition). (E and F) Naive splenic Smarta-1 transgenic CD4 + T cells were co-cultured with splenic DCs purified from WT or Il1rl1 −/− mice and 1 nM gp61 peptide with indicated cytokines and in the presence of PPARγ antagonist GW9662 or vehicle (DMSO) as a control. Frequencies of IL-5 + (E) and ST2 + (F) of CD4 + cells after 4 h restimulation with PMA/ionomycin ( n = 4/condition) for co-culture with) WT (E and Il1rl1 −/− (F) splenic DCs ( n = 4/condition). Data are means ± SEM and are representative of two independent experiments. The Student’s t test (unpaired) was used. *, P
Figure Legend Snippet: PPARγ suppresses a Th17/Th22 transcriptional program and is essential for IL-33–induced CD4 + T cell effector cytokine production. Naive splenic Smarta-1 transgenic CD4 + T cells were co-cultured with splenic DCs and 10 nM gp33 peptide in Th2 polarizing conditions in the presence of PPARγ antagonist GW9662 or vehicle (DMSO) as a control. After 4 d of co-culture, RNA sequencing of CD4 + T cells was performed ( n = 2). Heat map of the 100 most-differentially regulated genes (A) and expression levels of selected Th2 cytokines (B). (C) Naive splenic Smarta-1 transgenic CD4 + T cells were co-cultured with splenic DCs and 10 nM gp61 peptide in Th2 or Th22 polarizing conditions in the presence of PPARγ antagonist GW9662 or vehicle (DMSO) as a control. (C and D) Shown is the production of IL-17F (C) and IL-22 (D) after 4-h restimulation with PMA/ionomycin ( n = 4/condition). (E and F) Naive splenic Smarta-1 transgenic CD4 + T cells were co-cultured with splenic DCs purified from WT or Il1rl1 −/− mice and 1 nM gp61 peptide with indicated cytokines and in the presence of PPARγ antagonist GW9662 or vehicle (DMSO) as a control. Frequencies of IL-5 + (E) and ST2 + (F) of CD4 + cells after 4 h restimulation with PMA/ionomycin ( n = 4/condition) for co-culture with) WT (E and Il1rl1 −/− (F) splenic DCs ( n = 4/condition). Data are means ± SEM and are representative of two independent experiments. The Student’s t test (unpaired) was used. *, P

Techniques Used: Transgenic Assay, Cell Culture, Co-Culture Assay, RNA Sequencing Assay, Expressing, Purification, Mouse Assay

PPARγ is highly expressed in human effector memory Th2 cells. Effector memory T cells were sorted from PBMCs of human allergic and non-allergic donors, according to chemokine receptor expression and subsequently cultured in vitro as described in the Materials and methods section. T cells were pregated as CD4 + CD8 – CD14 – CD19 – CD25 – CD56 – CD45RA – CCR7 – cells, and subsequently, T cell subsets were identified as follows: CRTh2 + CCR4 + CXCR3 – CCR6 – (enriched in inflammatory Th2 cells); CCR4 + CRTh2 – CXCR3 – CCR6 – (enriched in Th2 cells); CXCR3 + CCR4 – CCR6 – (enriched in Th1 cells); CXCR3 + CCR6 + CCR4 – (enriched in Th1* cells); and CCR4 + CCR6 + CXCR3 – (enriched in Th17 cells). (A and B) Representative dot plots of the cytokine production of the different T cell clone types analyzed from an allergic donor after restimulation with PMA/ionomycin (A) and a summary of all clones tested for that donor (B). (C–F) mRNA expression of PPARG-1 and PPARG-2 of two allergic (IgE + ; C and D) and two non-allergic (IgE − ; E and F) donor-derived clones after restimulation with PMA/ionomycin.(G and H) mRNA expression of PPARG-1 (G) and PPARG-2 (H) of memory T cell subsets sorted directly ex vivo from PBMCs. Data are means ± SEM, and the sample size is visible in each graph. ANOVA (one-way) was used. *, P
Figure Legend Snippet: PPARγ is highly expressed in human effector memory Th2 cells. Effector memory T cells were sorted from PBMCs of human allergic and non-allergic donors, according to chemokine receptor expression and subsequently cultured in vitro as described in the Materials and methods section. T cells were pregated as CD4 + CD8 – CD14 – CD19 – CD25 – CD56 – CD45RA – CCR7 – cells, and subsequently, T cell subsets were identified as follows: CRTh2 + CCR4 + CXCR3 – CCR6 – (enriched in inflammatory Th2 cells); CCR4 + CRTh2 – CXCR3 – CCR6 – (enriched in Th2 cells); CXCR3 + CCR4 – CCR6 – (enriched in Th1 cells); CXCR3 + CCR6 + CCR4 – (enriched in Th1* cells); and CCR4 + CCR6 + CXCR3 – (enriched in Th17 cells). (A and B) Representative dot plots of the cytokine production of the different T cell clone types analyzed from an allergic donor after restimulation with PMA/ionomycin (A) and a summary of all clones tested for that donor (B). (C–F) mRNA expression of PPARG-1 and PPARG-2 of two allergic (IgE + ; C and D) and two non-allergic (IgE − ; E and F) donor-derived clones after restimulation with PMA/ionomycin.(G and H) mRNA expression of PPARG-1 (G) and PPARG-2 (H) of memory T cell subsets sorted directly ex vivo from PBMCs. Data are means ± SEM, and the sample size is visible in each graph. ANOVA (one-way) was used. *, P

Techniques Used: Expressing, Cell Culture, In Vitro, Clone Assay, Derivative Assay, Ex Vivo

PPARγ in T cells mediates development of pulmonary allergic inflammation. (A–I) Pparg fl/fl and Cd4 -Cre Pparg fl/fl mice were sensitized intratracheally with 10 µg HDM on d 0 and subsequently challenged intratracheally with 10 µg HDM or PBS on days 7–11. Animals were analyzed on day 14. Flow cytometry was used to characterize and quantitate BAL and lung cell populations, including eosinophils (A) and neutrophils (B). (B, top) Hematoxylin and eosin histology. (B, bottom) PAS and Alcian blue histology. CD4 + and CD8 + T cells (C) and BAL CD4 + T cells (D) were restimulated with PMA/ionomycin for 4 h. Shown is the frequency of CD4 + T cells that produced the indicated cytokines. The data presented are pooled from two independent experiments ( n = 5–11/group). Shown are ST2 + CD4 + Th2 cells (E), lin − CD90 + CD127 + CD25 + ILC2s (F), Foxp3 + CD4 + Treg cells (G), and GATA3 + and RORγt + cells (H) among CD4 + FoxP3 + Treg ( n = 4–6/group). (I) DEREG mice were treated with HDM as described in this legend. GFP + CD4 + Treg, ST2 + CD4 + Th2, and ST2 − CD4 + non-Th2 cells were sorted on day 14 by flow cytometry. Shown is the Pparg mRNA expression measured by quantitative PCR ( n = 3/group). The data are representative of three experiments. Data are means ± SEM and the sample size ( n ). The Student’s t test (unpaired) was used. *, P
Figure Legend Snippet: PPARγ in T cells mediates development of pulmonary allergic inflammation. (A–I) Pparg fl/fl and Cd4 -Cre Pparg fl/fl mice were sensitized intratracheally with 10 µg HDM on d 0 and subsequently challenged intratracheally with 10 µg HDM or PBS on days 7–11. Animals were analyzed on day 14. Flow cytometry was used to characterize and quantitate BAL and lung cell populations, including eosinophils (A) and neutrophils (B). (B, top) Hematoxylin and eosin histology. (B, bottom) PAS and Alcian blue histology. CD4 + and CD8 + T cells (C) and BAL CD4 + T cells (D) were restimulated with PMA/ionomycin for 4 h. Shown is the frequency of CD4 + T cells that produced the indicated cytokines. The data presented are pooled from two independent experiments ( n = 5–11/group). Shown are ST2 + CD4 + Th2 cells (E), lin − CD90 + CD127 + CD25 + ILC2s (F), Foxp3 + CD4 + Treg cells (G), and GATA3 + and RORγt + cells (H) among CD4 + FoxP3 + Treg ( n = 4–6/group). (I) DEREG mice were treated with HDM as described in this legend. GFP + CD4 + Treg, ST2 + CD4 + Th2, and ST2 − CD4 + non-Th2 cells were sorted on day 14 by flow cytometry. Shown is the Pparg mRNA expression measured by quantitative PCR ( n = 3/group). The data are representative of three experiments. Data are means ± SEM and the sample size ( n ). The Student’s t test (unpaired) was used. *, P

Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Produced, Expressing, Real-time Polymerase Chain Reaction

PPARγ in DCs modulates Th2 polarization in vivo . (A) The generation of Cd11c -Cre Pparg fl/fl mice containing WT AMs is shown schematically. Fetal (E17.5) lung AM precursors from WT CD45.1 + C57BL/6 embryos were transferred intranasally to neonatal (day 3 after birth) Pparg fl/fl and Cd11c -Cre Pparg fl/fl mice. After 8 wk, animals showed a reconstitution of Cd11c -Cre Pparg fl/fl mice with mature WT AMs, and they were subsequently used in HDM or OVA/alum asthma protocols, as described in Fig. 1 and Fig. S1. (B) Animals were sensitized with HDM and challenged with HDM or PBS with total cell numbers in the BAL (B) and lung (C) of eosinophils and neutrophils ( n = 4–8/group). (D, left) Hematoxylin and eosin (H E) histology. (D, right) PAS and Alcian blue histology. (E) Representative sections of total cell numbers of CD4 + and CD8 + T cells in the BAL. (F) BAL CD4 + T cells were restimulated with PMA/ionomycin for 4 h, and intracellular cytokine production was quantified, with the frequency of indicated cytokines ( n = 4–8/group). (G) Total number of lung ILC2s are shown. (H–J) WT and Cd11c -Cre Pparg fl/fl mice were sensitized with OVA/alum i.p. and challenged with OVA intratracheally (H). Shown are total cell numbers in the lung of eosinophils and neutrophils (I) and CD4 + and CD8 + T cells (J). (J) BAL CD4 + T cells were restimulated with PMA/ionomycin for 4 h, and intracellular cytokine production was quantified. Frequency of IFN-γ + , IL-17A + , and IL-4 + cells as well as the MFI for IL-4 ( n = 5/group). The data are representative of two experiments for each panel and are means ± SEM, with the sample size ( n ). The Student’s t test (unpaired) was used. *, P
Figure Legend Snippet: PPARγ in DCs modulates Th2 polarization in vivo . (A) The generation of Cd11c -Cre Pparg fl/fl mice containing WT AMs is shown schematically. Fetal (E17.5) lung AM precursors from WT CD45.1 + C57BL/6 embryos were transferred intranasally to neonatal (day 3 after birth) Pparg fl/fl and Cd11c -Cre Pparg fl/fl mice. After 8 wk, animals showed a reconstitution of Cd11c -Cre Pparg fl/fl mice with mature WT AMs, and they were subsequently used in HDM or OVA/alum asthma protocols, as described in Fig. 1 and Fig. S1. (B) Animals were sensitized with HDM and challenged with HDM or PBS with total cell numbers in the BAL (B) and lung (C) of eosinophils and neutrophils ( n = 4–8/group). (D, left) Hematoxylin and eosin (H E) histology. (D, right) PAS and Alcian blue histology. (E) Representative sections of total cell numbers of CD4 + and CD8 + T cells in the BAL. (F) BAL CD4 + T cells were restimulated with PMA/ionomycin for 4 h, and intracellular cytokine production was quantified, with the frequency of indicated cytokines ( n = 4–8/group). (G) Total number of lung ILC2s are shown. (H–J) WT and Cd11c -Cre Pparg fl/fl mice were sensitized with OVA/alum i.p. and challenged with OVA intratracheally (H). Shown are total cell numbers in the lung of eosinophils and neutrophils (I) and CD4 + and CD8 + T cells (J). (J) BAL CD4 + T cells were restimulated with PMA/ionomycin for 4 h, and intracellular cytokine production was quantified. Frequency of IFN-γ + , IL-17A + , and IL-4 + cells as well as the MFI for IL-4 ( n = 5/group). The data are representative of two experiments for each panel and are means ± SEM, with the sample size ( n ). The Student’s t test (unpaired) was used. *, P

Techniques Used: In Vivo, Mouse Assay, Affinity Magnetic Separation

71) Product Images from "Mycobacterium tuberculosis Activates Human Macrophage Peroxisome Proliferator-Activated Receptor ? Linking Mannose Receptor Recognition to Regulation of Immune Responses"

Article Title: Mycobacterium tuberculosis Activates Human Macrophage Peroxisome Proliferator-Activated Receptor ? Linking Mannose Receptor Recognition to Regulation of Immune Responses

Journal: Journal of immunology (Baltimore, Md. : 1950)

doi: 10.4049/jimmunol.1000866

PPARγ knockdown and modulation of activity alter ManLAM-mediated IL-8 release. MDMs were transfected with scramble siRNA (control) or PPARγ siRNA by nucleofection. After 16 h, cells were stimulated with ManLAM (5 μg/ml) for 6 and
Figure Legend Snippet: PPARγ knockdown and modulation of activity alter ManLAM-mediated IL-8 release. MDMs were transfected with scramble siRNA (control) or PPARγ siRNA by nucleofection. After 16 h, cells were stimulated with ManLAM (5 μg/ml) for 6 and

Techniques Used: Activity Assay, Transfection

M. bovis BCG infection induces limited PPARγ expression in macrophages. A , MDMs were infected with M. tuberculosis H 37 R v or BCG at an MOI of 5:1. After 2 h, the cells were washed and incubated in 2% autologous serum for 6, 24, 48, and 72 h. Cell
Figure Legend Snippet: M. bovis BCG infection induces limited PPARγ expression in macrophages. A , MDMs were infected with M. tuberculosis H 37 R v or BCG at an MOI of 5:1. After 2 h, the cells were washed and incubated in 2% autologous serum for 6, 24, 48, and 72 h. Cell

Techniques Used: Infection, Expressing, Incubation

Model of the PPARγ signaling pathway in human macrophages in response to virulent M. tuberculosis or ManLAM. Shown is a schematic based on our results along with those in the literature of how virulent M. tuberculosis infection or ManLAM stimulation
Figure Legend Snippet: Model of the PPARγ signaling pathway in human macrophages in response to virulent M. tuberculosis or ManLAM. Shown is a schematic based on our results along with those in the literature of how virulent M. tuberculosis infection or ManLAM stimulation

Techniques Used: Infection

The macrophage MR regulates PPARγ expression in response to ManLAM. MDMs were transfected with MR siRNA or scramble siRNA by nucleofection and plated in RPMI 1640 containing 20% autologous serum. After 48 h, the cells were washed and stimulated
Figure Legend Snippet: The macrophage MR regulates PPARγ expression in response to ManLAM. MDMs were transfected with MR siRNA or scramble siRNA by nucleofection and plated in RPMI 1640 containing 20% autologous serum. After 48 h, the cells were washed and stimulated

Techniques Used: Expressing, Transfection

M. tuberculosis H 37 R v and its cell wall component ManLAM upregulate PPARγ expression in human macrophages. MDM monolayers were incubated with M. tuberculosis H 37 R v (MOI 5:1) ( A ) for 2 h; cells were washed and incubated in 2% autologous serum for
Figure Legend Snippet: M. tuberculosis H 37 R v and its cell wall component ManLAM upregulate PPARγ expression in human macrophages. MDM monolayers were incubated with M. tuberculosis H 37 R v (MOI 5:1) ( A ) for 2 h; cells were washed and incubated in 2% autologous serum for

Techniques Used: Expressing, Incubation

PPARγ knockdown leads to decreased growth of M. tuberculosis in human macrophages and increased TNF production. A , MDMs were transfected with scrambled siRNA or PPARγ siRNA by nucleofection. After 24 h, the cells were incubated with M.
Figure Legend Snippet: PPARγ knockdown leads to decreased growth of M. tuberculosis in human macrophages and increased TNF production. A , MDMs were transfected with scrambled siRNA or PPARγ siRNA by nucleofection. After 24 h, the cells were incubated with M.

Techniques Used: Transfection, Incubation

ManLAM activation of MAPK-p38 regulates the PPARγ-mediated IL-8 response through activation of cPLA 2 in macrophages. MDMs were pretreated with the p38 inhibitor SB-203580 (5 μM) or DMSO for 30 min and subsequently stimulated with ManLAM
Figure Legend Snippet: ManLAM activation of MAPK-p38 regulates the PPARγ-mediated IL-8 response through activation of cPLA 2 in macrophages. MDMs were pretreated with the p38 inhibitor SB-203580 (5 μM) or DMSO for 30 min and subsequently stimulated with ManLAM

Techniques Used: Activation Assay

72) Product Images from "CD44 Loss Disrupts Lung Lipid Surfactant Homeostasis and Exacerbates Oxidized Lipid-Induced Lung Inflammation"

Article Title: CD44 Loss Disrupts Lung Lipid Surfactant Homeostasis and Exacerbates Oxidized Lipid-Induced Lung Inflammation

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2020.00029

PPARγ expression is defective in CD44 −/− AMs. (A) Representative confocal microscopy showing CD44 +/+ and CD44 −/− AMs labeled with intracellular PPARγ antibody and DAPI. (B) Comparison of nuclear PPARγ mean pixel intensity (MPI) between CD44 +/+ and CD44 −/− AMs, determined by confocal microscopy. (C) Representative flow cytometry histograms and graphs comparing intracellular PPARγ expression between CD44 +/+ and CD44 −/− AMs. (D,E) Representative flow cytometry histograms and graphs comparing the levels of CD36, BODIPY and CD11c levels by MFI (after subtraction of background autofluorescence) in CD44 +/+ AMs cultured with CSF-2 or CSF-2 and the PPARγ antagonist T0070907 (Antag) for 48 h in culture. Data show an average of three to five mice from each experiment ± SD, repeated twice or three times; for confocal imaging three to four fields containing cells were analyzed per mice. Significance indicated as * p
Figure Legend Snippet: PPARγ expression is defective in CD44 −/− AMs. (A) Representative confocal microscopy showing CD44 +/+ and CD44 −/− AMs labeled with intracellular PPARγ antibody and DAPI. (B) Comparison of nuclear PPARγ mean pixel intensity (MPI) between CD44 +/+ and CD44 −/− AMs, determined by confocal microscopy. (C) Representative flow cytometry histograms and graphs comparing intracellular PPARγ expression between CD44 +/+ and CD44 −/− AMs. (D,E) Representative flow cytometry histograms and graphs comparing the levels of CD36, BODIPY and CD11c levels by MFI (after subtraction of background autofluorescence) in CD44 +/+ AMs cultured with CSF-2 or CSF-2 and the PPARγ antagonist T0070907 (Antag) for 48 h in culture. Data show an average of three to five mice from each experiment ± SD, repeated twice or three times; for confocal imaging three to four fields containing cells were analyzed per mice. Significance indicated as * p

Techniques Used: Expressing, Affinity Magnetic Separation, Confocal Microscopy, Labeling, Flow Cytometry, Cell Culture, Mouse Assay, Imaging

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Luciferase:

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Article Snippet: Reagents and antibodies RPMI and DMEM, DMEM F12 tissue culture media, Lipofectamine 2000 and β-galactosidase assay kit were purchased from Invitrogen; the luciferase Assay Reagent from Promega (Madison, WI); Troglitazone, TRAIL and Cycloheximide (CHX) were purchased from EMD Biosciences (Gibbstown, NJ), Compound C, AKT Inhibitor VIII, LY294002, Kenpaullone and AR-A014418 were from EMD Millipore (Billerica, MA), CHIR 99021 was from Sigma (St. Louis, MO). .. The antibodies utilized were obtained from the following sources: poly (ADP-ribose) polymerase (PARP), caspase-3, GSK-3β, phospho-GSK-3βSer9 , GSK3α, AKT, pAKTSer473 , pAKT2Ser474 , PPARγ, AMPKα1 and α2, GS, pGSSer641 from Cell Signaling Technologies (Danvers, MA); GAPDH from Ambion Inc. (Austin, TX).

Blocking Assay:

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Incubation:

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Activity Assay:

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Expressing:

Article Title: Prokineticin receptor-1-dependent paracrine and autocrine pathways control cardiac tcf21+ fibroblast progenitor cell transformation into adipocytes and vascular cells
Article Snippet: The blots were incubated with a blocking solution and then incubated overnight with primary antibodies against peroxisome proliferator-activated receptor gamma (PPARγ, Cell Signaling Technology) at 4 °C with gentle shaking with primary antibodies to PPARγ (cell Signaling Technology). .. The signals were quantified by scanning laser densitometry and normalized to total amounts of the corresponding reference protein expression levels.

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Article Snippet: The membrane was probed with rabbit anti-PPARα (Assay Biotech), PPARβ (Abcam), and PPARγ (Cell Signaling) antibody (R & D System). .. Protein loading was normalized to GAPDH expression using anti-mouse GAPDH antibody (Sigma-Aldrich, St. Louis, MO).

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Article Snippet: Western blot analysis PPARγ and NF-κB protein expression were examined using western blot analysis. .. The supernatant fractions were assayed for protein concentration using a Bradford reagent (Bio-Rad, Richmond, CA, USA) and were used for the western blot analyses of PPARγ, NF-κB, and β-actin (Cell Signaling, Beverly, MA, USA).

Western Blot:

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Article Title: Neonatal diethylstilbestrol exposure alters the metabolic profile of uterine epithelial cells
Article Snippet: .. Antibodies and dilutions used were: 1:100 (immunohistochemistry) and 1:1000 (western blot) for PPARγ (Cell Signaling); 1:5000 (western blot) for GAPDH (Cell Signaling); 1:200 (immunofluorescence) for KLF4 (Santa Cruz Biotechnology); 1:100 (immunofluorescence) for G6PD and GLUT1 (generous gifts from Kelle Moley, Washington University, St Louis, MO); 1:100 for HK2 (Cell Signaling); 1:1000 for Alexa-Fluor-594-conjugated goat anti-rabbit (Invitrogen) and 1:5000 for HRP-conjugated goat anti-rabbit secondary antibodies. .. For ORO staining, tissues were fixed in 4% paraformaldehyde briefly, washed through a series of Tissue-Tek optimal cutting temperature compound (OCT)-sucrose solutions, and embedded in OCT Slides of 10-μm frozen sections were washed in water once, twice in 100% propylene glycol (Sigma) and then stained in 0.7% ORO in propylene glycol for 7 minutes with agitation at 60°C.

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Article Title: Licochalcone E protects against carbon tetrachloride-induced liver toxicity by activating peroxisome proliferator-activated receptor gamma
Article Snippet: .. The supernatant fractions were assayed for protein concentration using a Bradford reagent (Bio-Rad, Richmond, CA, USA) and were used for the western blot analyses of PPARγ, NF-κB, and β-actin (Cell Signaling, Beverly, MA, USA). .. Horseradish peroxidase-conjugated IgG (Zymed, South San Francisco, CA, USA) was used as a secondary antibody.

Immunohistochemistry:

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Protease Inhibitor:

Article Title: Peroxisome Proliferator-Activated Receptor γ Expression Is Inversely Associated with Macroscopic Vascular Invasion in Human Hepatocellular Carcinoma
Article Snippet: Western Blot Analyses Total proteins were extracted from cells using RIPA buffer (150 mM NaCl, 50 mM Tris-HCl, 0.25% sodium deoxycholate, 1% Triton X-100, 0.1% SDS), supplemented with a protease inhibitor cocktail (Calbiochem, San Diego, CA, USA). .. PPARγ (Cell Signaling, Danvers, MA, USA), E-cadherin (Cell Signaling), signal transducer and activator transcription 3 (STAT3) (Cell Signaling), cyclin D1 (Millipore, Darmstadt, Germany), and β-actin (Sigma-Aldrich) antibodies were used to probe the proteins on the membrane at 4 °C overnight.

Cell Culture:

Article Title: Smooth Muscle Proliferation and Role of the Prostacyclin (IP) Receptor in Idiopathic Pulmonary Arterial Hypertension
Article Snippet: Blood vessels were immunostained as previously described ( ) using cell-specific markers (Figure E2C and E2D) and antibodies to PPARγ (Cell Signaling Technology, Danvers MA) and the IP receptor. .. For immunofluoresence, monolayers of cultured cells were fixed and permeabilized and then stained for the IP receptor and nuclei (TO-PRO-3; Invitrogen, Paisley, UK).

Imaging:

Article Title: Licochalcone E protects against carbon tetrachloride-induced liver toxicity by activating peroxisome proliferator-activated receptor gamma
Article Snippet: The supernatant fractions were assayed for protein concentration using a Bradford reagent (Bio-Rad, Richmond, CA, USA) and were used for the western blot analyses of PPARγ, NF-κB, and β-actin (Cell Signaling, Beverly, MA, USA). .. Finally, the bands were visualized using ECL-plus reagent, and the Bio-Rad Gel Doc 2000 imaging system and software were used to calculate the integrated absorbance (IA) of the bands.

Protein Concentration:

Article Title: Licochalcone E protects against carbon tetrachloride-induced liver toxicity by activating peroxisome proliferator-activated receptor gamma
Article Snippet: .. The supernatant fractions were assayed for protein concentration using a Bradford reagent (Bio-Rad, Richmond, CA, USA) and were used for the western blot analyses of PPARγ, NF-κB, and β-actin (Cell Signaling, Beverly, MA, USA). .. Horseradish peroxidase-conjugated IgG (Zymed, South San Francisco, CA, USA) was used as a secondary antibody.

Recombinant:

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Article Snippet: Reagents Recombinant human BMPs: BMP6 (507-BP-020), BMP7 (354-BP-010) and BMP8 (1073-BP-010) were purchased from R & D Systems. .. The following antibodies were purchased from Santa Cruz Biotechnology: Optn (sc-166576), Gapdh (sc-32233) and the following were purchased from Cell Signaling: Pparγ (2435S), Fabp4 (3544S), Adiponectin (2789S), and Cox2 (12282S).

Immunofluorescence:

Article Title: Neonatal diethylstilbestrol exposure alters the metabolic profile of uterine epithelial cells
Article Snippet: .. Antibodies and dilutions used were: 1:100 (immunohistochemistry) and 1:1000 (western blot) for PPARγ (Cell Signaling); 1:5000 (western blot) for GAPDH (Cell Signaling); 1:200 (immunofluorescence) for KLF4 (Santa Cruz Biotechnology); 1:100 (immunofluorescence) for G6PD and GLUT1 (generous gifts from Kelle Moley, Washington University, St Louis, MO); 1:100 for HK2 (Cell Signaling); 1:1000 for Alexa-Fluor-594-conjugated goat anti-rabbit (Invitrogen) and 1:5000 for HRP-conjugated goat anti-rabbit secondary antibodies. .. For ORO staining, tissues were fixed in 4% paraformaldehyde briefly, washed through a series of Tissue-Tek optimal cutting temperature compound (OCT)-sucrose solutions, and embedded in OCT Slides of 10-μm frozen sections were washed in water once, twice in 100% propylene glycol (Sigma) and then stained in 0.7% ORO in propylene glycol for 7 minutes with agitation at 60°C.

Article Title: Smooth Muscle Proliferation and Role of the Prostacyclin (IP) Receptor in Idiopathic Pulmonary Arterial Hypertension
Article Snippet: Paragraph title: Immunohistochemistry and Immunofluorescence ... Blood vessels were immunostained as previously described ( ) using cell-specific markers (Figure E2C and E2D) and antibodies to PPARγ (Cell Signaling Technology, Danvers MA) and the IP receptor.

Microscopy:

Article Title: Smooth Muscle Proliferation and Role of the Prostacyclin (IP) Receptor in Idiopathic Pulmonary Arterial Hypertension
Article Snippet: Blood vessels were immunostained as previously described ( ) using cell-specific markers (Figure E2C and E2D) and antibodies to PPARγ (Cell Signaling Technology, Danvers MA) and the IP receptor. .. Slides were examined using a Leica DM LB microscope (Leica Microsystems, Wetzlar, Germany), and images were acquired and analyzed in a blinded fashion (details provided in the online supplement).

Protein Extraction:

Article Title: Netrin-1-treated macrophages protect the kidney against ischemia-reperfusion injury and suppress inflammation by inducing M2 polarization
Article Snippet: Protein extraction from RAW264.7 cells and Western blot analysis were carried out as described previously ( , ). .. The membrane was probed with rabbit anti-PPARα (Assay Biotech), PPARβ (Abcam), and PPARγ (Cell Signaling) antibody (R & D System).

Article Title: Resolvin D1 attenuates inflammation in lipopolysaccharide-induced acute lung injury through a process involving the PPAR?/NF-?B pathway
Article Snippet: Western Blot analysis Lung tissue samples collected at 6 h after LPS treatment were homogenized, and cytoplasmic and nuclear proteins were extracted separately using the Nuclear and Cytoplasmic Protein Extraction Kit (Viagene Biotech, Ningbo, China) according to the manufacturer's instructions. .. Mouse polyclonal antibodies were used against IκBα, the NF-κB p65 subunit, and PPARγ (Cell Signaling Technology, USA); histone H3.1 (Signalway Antibody, USA); and β-actin (Santa Cruz Biotechnology, USA).

Lysis:

Article Title: Licochalcone E protects against carbon tetrachloride-induced liver toxicity by activating peroxisome proliferator-activated receptor gamma
Article Snippet: The protein extracts from the liver tissue were prepared using a lysis buffer (50 mM Tris-HCl, pH 7.6, 0.5% Triton X-100, and 20% glycerol). .. The supernatant fractions were assayed for protein concentration using a Bradford reagent (Bio-Rad, Richmond, CA, USA) and were used for the western blot analyses of PPARγ, NF-κB, and β-actin (Cell Signaling, Beverly, MA, USA).

IA:

Article Title: Licochalcone E protects against carbon tetrachloride-induced liver toxicity by activating peroxisome proliferator-activated receptor gamma
Article Snippet: The supernatant fractions were assayed for protein concentration using a Bradford reagent (Bio-Rad, Richmond, CA, USA) and were used for the western blot analyses of PPARγ, NF-κB, and β-actin (Cell Signaling, Beverly, MA, USA). .. Finally, the bands were visualized using ECL-plus reagent, and the Bio-Rad Gel Doc 2000 imaging system and software were used to calculate the integrated absorbance (IA) of the bands.

SDS Page:

Article Title: Prokineticin receptor-1-dependent paracrine and autocrine pathways control cardiac tcf21+ fibroblast progenitor cell transformation into adipocytes and vascular cells
Article Snippet: The proteins were separated under denaturing conditions by SDS-PAGE (10% gel) and transferred to a polyvinylidene difluoride (PVDF) membrane. .. The blots were incubated with a blocking solution and then incubated overnight with primary antibodies against peroxisome proliferator-activated receptor gamma (PPARγ, Cell Signaling Technology) at 4 °C with gentle shaking with primary antibodies to PPARγ (cell Signaling Technology).

Article Title: Peroxisome Proliferator-Activated Receptor γ Expression Is Inversely Associated with Macroscopic Vascular Invasion in Human Hepatocellular Carcinoma
Article Snippet: Proteins were separated via 10% SDS-PAGE gel and electrotransferred onto a polyvinylidene difluoride (PVDF) membrane. .. PPARγ (Cell Signaling, Danvers, MA, USA), E-cadherin (Cell Signaling), signal transducer and activator transcription 3 (STAT3) (Cell Signaling), cyclin D1 (Millipore, Darmstadt, Germany), and β-actin (Sigma-Aldrich) antibodies were used to probe the proteins on the membrane at 4 °C overnight.

Plasmid Preparation:

Article Title: Modulation of glycogen synthase kinase-3β following TRAIL combinatorial treatment in cancer cells
Article Snippet: The antibodies utilized were obtained from the following sources: poly (ADP-ribose) polymerase (PARP), caspase-3, GSK-3β, phospho-GSK-3βSer9 , GSK3α, AKT, pAKTSer473 , pAKT2Ser474 , PPARγ, AMPKα1 and α2, GS, pGSSer641 from Cell Signaling Technologies (Danvers, MA); GAPDH from Ambion Inc. (Austin, TX). .. The GSK3β promoter luciferase plasmid (pGL3-GSK-3β-luc (−427/+66) was obtained from the laboratory of Dr. Daniel D. Billadeau, Mayo Clinic, Rochester, MN [ ].

Article Title: mTOR Regulates Fatty Infiltration through SREBP-1 and PPARg after a Combined Massive Rotator Cuff Tear and Suprascapular Nerve Injury in Rats
Article Snippet: To localize p-mTOR, SREBP-1, and PPARγ activity within surgical and sham supraspinatus muscles, immunohistochemistry (IHC) was performed using phospho-mTOR, PPARγ, (Cell Signaling Technology, Inc.) and SREBP-1 (Novux Biologicals, Littleton, CO) antibodies at a dilution of 1:200. .. A DAB staining kit (Vector Laboratories, Inc., Burlingame, CA) was used for developing as previously described.

Software:

Article Title: Licochalcone E protects against carbon tetrachloride-induced liver toxicity by activating peroxisome proliferator-activated receptor gamma
Article Snippet: The supernatant fractions were assayed for protein concentration using a Bradford reagent (Bio-Rad, Richmond, CA, USA) and were used for the western blot analyses of PPARγ, NF-κB, and β-actin (Cell Signaling, Beverly, MA, USA). .. Finally, the bands were visualized using ECL-plus reagent, and the Bio-Rad Gel Doc 2000 imaging system and software were used to calculate the integrated absorbance (IA) of the bands.

Immunoprecipitation:

Article Title: TLE3 is a dual function transcriptional coregulator of adipogenesis
Article Snippet: For immunoprecipitation, nuclear extracts were diluted in IP buffer (20 mM Tris, 137 mM NaCl, 2 mM EDTA, 1% NP-40, 10% glycerol) and pre-cleared with ProteinA agarose beads (Santa Cruz). .. Extracts were mixed with IgG (PP64, Millipore), TCF4 (C48H11, Cell signaling) or PPARγ (81B8, Cell signaling) antibodies and incubated with beads.

Staining:

Article Title: Neonatal diethylstilbestrol exposure alters the metabolic profile of uterine epithelial cells
Article Snippet: Paragraph title: Western blot, immunohistochemistry, immunofluorescence and Oil-Red O staining ... Antibodies and dilutions used were: 1:100 (immunohistochemistry) and 1:1000 (western blot) for PPARγ (Cell Signaling); 1:5000 (western blot) for GAPDH (Cell Signaling); 1:200 (immunofluorescence) for KLF4 (Santa Cruz Biotechnology); 1:100 (immunofluorescence) for G6PD and GLUT1 (generous gifts from Kelle Moley, Washington University, St Louis, MO); 1:100 for HK2 (Cell Signaling); 1:1000 for Alexa-Fluor-594-conjugated goat anti-rabbit (Invitrogen) and 1:5000 for HRP-conjugated goat anti-rabbit secondary antibodies.

Article Title: Smooth Muscle Proliferation and Role of the Prostacyclin (IP) Receptor in Idiopathic Pulmonary Arterial Hypertension
Article Snippet: Blood vessels were immunostained as previously described ( ) using cell-specific markers (Figure E2C and E2D) and antibodies to PPARγ (Cell Signaling Technology, Danvers MA) and the IP receptor. .. For immunofluoresence, monolayers of cultured cells were fixed and permeabilized and then stained for the IP receptor and nuclei (TO-PRO-3; Invitrogen, Paisley, UK).

Article Title: mTOR Regulates Fatty Infiltration through SREBP-1 and PPARg after a Combined Massive Rotator Cuff Tear and Suprascapular Nerve Injury in Rats
Article Snippet: To localize p-mTOR, SREBP-1, and PPARγ activity within surgical and sham supraspinatus muscles, immunohistochemistry (IHC) was performed using phospho-mTOR, PPARγ, (Cell Signaling Technology, Inc.) and SREBP-1 (Novux Biologicals, Littleton, CO) antibodies at a dilution of 1:200. .. A DAB staining kit (Vector Laboratories, Inc., Burlingame, CA) was used for developing as previously described.

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    Cell Signaling Technology Inc antibodies against mouse pparγ
    ASA treatment promoted immunomodulatory properties of BMMSCs via the 15d-PGJ 2 <t>/PPARγ/TGF-β1</t> pathway. (A) Following ASA treatment (200 μg/mL), the addition of 15d-PGJ 2 (10 μmol) significantly suppressed the secretion of TGF-β1 by BMMSCs. (B) Anti-TGF-β1-neutralizing antibody (TGF-β1 Ab, 1 μg/mL) had no effect on 15d-PGJ 2 production. (C) ASA treatment (200 μg/mL) significantly increased the concentration of TGF-β1. (D) ASA treatment decreased the protein expression of nuclear PPARγ, whereas the protein levels of total PPARγ remained unchanged. (E) In the presence of ASA, rosiglitazone or 15d-PGJ 2 promoted PPARγ nuclear translocation, whereas GW9662 treatment blocked PPARγ nuclear translocation. (F) The upregulation of TGF-β1 after ASA treatment could be blocked by agonists of PPARγ, rosiglitazone (10 μmol) or 15d-PGJ 2 (10 μmol), whereas this blockage could be reversed by the pretreatment of GW9662 (1 μmol), an antagonist of PPARγ. (G) Schematic diagram indicating that ASA could promote TGF-β1 secretion of BMMSCs via the 15d-PGJ 2 /PPARγ/TGF-β1 pathway. AA, Arachidonic acid; COX-2, cyclooxygenase-2. The results are representative of at least three independent experiments. Results were expressed as mean±standard deviation (SD), and statistical significance was shown as * P
    Antibodies Against Mouse Pparγ, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc primary antibody ppar γ
    Effect of Portulaca extracts on NF- κ B and <t>PPAR-</t> γ expression in colonic tissue. Panel (a) represents the immunohistochemistry staining of PPAR- γ . (NC refers to negative control; DSS refers to DSS alone group; M refers to mesalamine group; P refers to Portulaca group, original magnification: ×200). Panel (b) shows the expression of PPAR- γ , NF- κ B, and pNF- κ B in colorectum tissues from western blot. Panel (c) shows the histograms of grey intensity for PPAR- γ and NF- κ B relative to β -actin. Panel (d) shows the relative mRNA expression of PPAR- γ and NF- κ B from quantitative real-time PCR. (Data are presented as mean ± SD from six experiments. ∗ indicates comparison between DSS and NC; # indicates comparison between mesalamine and DSS; + indicates the comparison between Portulaca and DSS. The number of symbols indicates significance of difference; for example, four symbols indicate p
    Primary Antibody Ppar γ, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit anti pparγ
    Role of MuRF2 in regulating PPAR isoform activity and its role in high fat diet cardiac hypertrophy in vivo. Isolation of cardiac nuclei from MuRF2−/− and sibling wild type mouse hearts revealed increases in a PPAR∝, PPARβ/δ, and <t>PPARγ</t> DNA binding activity using PPRE-DNA as bait and ELISA detection of PPARα protein (N = 4/group). b Experimental design of high fat diet (60%)-induced cardiomyopathy. c High fat diet induces cardiac MuRF2 levels after 26 weeks HFD (N = 3/group). d Endogenous MuRF2 inhibits HFD-induced LV Mass and heart wet weights, as MuRF2−/− hearts have a significant increase in heart weight normalized to body weight and tibia length (N = 5/group). e Endogenous MuRF2, found in skeletal muscle and the heart does not affect overall body weight (N indicated below graph). Values expressed as Mean ± SE. Statistical analysis was performed using a Student’s t-test comparing MuRF2−/− and MuRF2+/+ groups. *p ≤ 0.001, **p
    Rabbit Anti Pparγ, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc ppar γ
    (A) Effects of S -allyl cysteine (SAC) administration on the gene expression and protein levels of Peroxisome proliferators-activated receptor <t>(PPAR)-γ</t> in the liver. Animals were treated as described in Fig. 1 . PPAR-γ mRNA expression was measured as described in Methods. Values are means ± SEM ( n = 5–6). # p
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    ASA treatment promoted immunomodulatory properties of BMMSCs via the 15d-PGJ 2 /PPARγ/TGF-β1 pathway. (A) Following ASA treatment (200 μg/mL), the addition of 15d-PGJ 2 (10 μmol) significantly suppressed the secretion of TGF-β1 by BMMSCs. (B) Anti-TGF-β1-neutralizing antibody (TGF-β1 Ab, 1 μg/mL) had no effect on 15d-PGJ 2 production. (C) ASA treatment (200 μg/mL) significantly increased the concentration of TGF-β1. (D) ASA treatment decreased the protein expression of nuclear PPARγ, whereas the protein levels of total PPARγ remained unchanged. (E) In the presence of ASA, rosiglitazone or 15d-PGJ 2 promoted PPARγ nuclear translocation, whereas GW9662 treatment blocked PPARγ nuclear translocation. (F) The upregulation of TGF-β1 after ASA treatment could be blocked by agonists of PPARγ, rosiglitazone (10 μmol) or 15d-PGJ 2 (10 μmol), whereas this blockage could be reversed by the pretreatment of GW9662 (1 μmol), an antagonist of PPARγ. (G) Schematic diagram indicating that ASA could promote TGF-β1 secretion of BMMSCs via the 15d-PGJ 2 /PPARγ/TGF-β1 pathway. AA, Arachidonic acid; COX-2, cyclooxygenase-2. The results are representative of at least three independent experiments. Results were expressed as mean±standard deviation (SD), and statistical significance was shown as * P

    Journal: Stem Cells and Development

    Article Title: Aspirin Treatment Improved Mesenchymal Stem Cell Immunomodulatory Properties via the 15d-PGJ2/PPARγ/TGF-β1 Pathway

    doi: 10.1089/scd.2014.0081

    Figure Lengend Snippet: ASA treatment promoted immunomodulatory properties of BMMSCs via the 15d-PGJ 2 /PPARγ/TGF-β1 pathway. (A) Following ASA treatment (200 μg/mL), the addition of 15d-PGJ 2 (10 μmol) significantly suppressed the secretion of TGF-β1 by BMMSCs. (B) Anti-TGF-β1-neutralizing antibody (TGF-β1 Ab, 1 μg/mL) had no effect on 15d-PGJ 2 production. (C) ASA treatment (200 μg/mL) significantly increased the concentration of TGF-β1. (D) ASA treatment decreased the protein expression of nuclear PPARγ, whereas the protein levels of total PPARγ remained unchanged. (E) In the presence of ASA, rosiglitazone or 15d-PGJ 2 promoted PPARγ nuclear translocation, whereas GW9662 treatment blocked PPARγ nuclear translocation. (F) The upregulation of TGF-β1 after ASA treatment could be blocked by agonists of PPARγ, rosiglitazone (10 μmol) or 15d-PGJ 2 (10 μmol), whereas this blockage could be reversed by the pretreatment of GW9662 (1 μmol), an antagonist of PPARγ. (G) Schematic diagram indicating that ASA could promote TGF-β1 secretion of BMMSCs via the 15d-PGJ 2 /PPARγ/TGF-β1 pathway. AA, Arachidonic acid; COX-2, cyclooxygenase-2. The results are representative of at least three independent experiments. Results were expressed as mean±standard deviation (SD), and statistical significance was shown as * P

    Article Snippet: After blocking with TBS/5% nonfat dry milk (Applygen) for 1 h, the membrane was incubated with antibodies against mouse PPARγ (Cell Signaling Technology), Histone H1 (BioWorld), and β-actin (Sigma) overnight at 4°C.

    Techniques: Concentration Assay, Expressing, Translocation Assay, Standard Deviation

    Effect of Portulaca extracts on NF- κ B and PPAR- γ expression in colonic tissue. Panel (a) represents the immunohistochemistry staining of PPAR- γ . (NC refers to negative control; DSS refers to DSS alone group; M refers to mesalamine group; P refers to Portulaca group, original magnification: ×200). Panel (b) shows the expression of PPAR- γ , NF- κ B, and pNF- κ B in colorectum tissues from western blot. Panel (c) shows the histograms of grey intensity for PPAR- γ and NF- κ B relative to β -actin. Panel (d) shows the relative mRNA expression of PPAR- γ and NF- κ B from quantitative real-time PCR. (Data are presented as mean ± SD from six experiments. ∗ indicates comparison between DSS and NC; # indicates comparison between mesalamine and DSS; + indicates the comparison between Portulaca and DSS. The number of symbols indicates significance of difference; for example, four symbols indicate p

    Journal: PPAR Research

    Article Title: Portulaca Extract Attenuates Development of Dextran Sulfate Sodium Induced Colitis in Mice through Activation of PPARγ

    doi: 10.1155/2018/6079101

    Figure Lengend Snippet: Effect of Portulaca extracts on NF- κ B and PPAR- γ expression in colonic tissue. Panel (a) represents the immunohistochemistry staining of PPAR- γ . (NC refers to negative control; DSS refers to DSS alone group; M refers to mesalamine group; P refers to Portulaca group, original magnification: ×200). Panel (b) shows the expression of PPAR- γ , NF- κ B, and pNF- κ B in colorectum tissues from western blot. Panel (c) shows the histograms of grey intensity for PPAR- γ and NF- κ B relative to β -actin. Panel (d) shows the relative mRNA expression of PPAR- γ and NF- κ B from quantitative real-time PCR. (Data are presented as mean ± SD from six experiments. ∗ indicates comparison between DSS and NC; # indicates comparison between mesalamine and DSS; + indicates the comparison between Portulaca and DSS. The number of symbols indicates significance of difference; for example, four symbols indicate p

    Article Snippet: Primary antibody PPAR- γ (Cell Signaling Technology, Danvers, MA, USA) was diluted into working concentration and then incubated with the slices at 4°C overnight.

    Techniques: Expressing, Immunohistochemistry, Staining, Negative Control, Western Blot, Real-time Polymerase Chain Reaction

    Role of MuRF2 in regulating PPAR isoform activity and its role in high fat diet cardiac hypertrophy in vivo. Isolation of cardiac nuclei from MuRF2−/− and sibling wild type mouse hearts revealed increases in a PPAR∝, PPARβ/δ, and PPARγ DNA binding activity using PPRE-DNA as bait and ELISA detection of PPARα protein (N = 4/group). b Experimental design of high fat diet (60%)-induced cardiomyopathy. c High fat diet induces cardiac MuRF2 levels after 26 weeks HFD (N = 3/group). d Endogenous MuRF2 inhibits HFD-induced LV Mass and heart wet weights, as MuRF2−/− hearts have a significant increase in heart weight normalized to body weight and tibia length (N = 5/group). e Endogenous MuRF2, found in skeletal muscle and the heart does not affect overall body weight (N indicated below graph). Values expressed as Mean ± SE. Statistical analysis was performed using a Student’s t-test comparing MuRF2−/− and MuRF2+/+ groups. *p ≤ 0.001, **p

    Journal: Cardiovascular Diabetology

    Article Title: MuRF2 regulates PPARγ1 activity to protect against diabetic cardiomyopathy and enhance weight gain induced by a high fat diet

    doi: 10.1186/s12933-015-0252-x

    Figure Lengend Snippet: Role of MuRF2 in regulating PPAR isoform activity and its role in high fat diet cardiac hypertrophy in vivo. Isolation of cardiac nuclei from MuRF2−/− and sibling wild type mouse hearts revealed increases in a PPAR∝, PPARβ/δ, and PPARγ DNA binding activity using PPRE-DNA as bait and ELISA detection of PPARα protein (N = 4/group). b Experimental design of high fat diet (60%)-induced cardiomyopathy. c High fat diet induces cardiac MuRF2 levels after 26 weeks HFD (N = 3/group). d Endogenous MuRF2 inhibits HFD-induced LV Mass and heart wet weights, as MuRF2−/− hearts have a significant increase in heart weight normalized to body weight and tibia length (N = 5/group). e Endogenous MuRF2, found in skeletal muscle and the heart does not affect overall body weight (N indicated below graph). Values expressed as Mean ± SE. Statistical analysis was performed using a Student’s t-test comparing MuRF2−/− and MuRF2+/+ groups. *p ≤ 0.001, **p

    Article Snippet: Rabbit anti-PPARα (Abcam Inc. Cat. #24509,1:1000), rabbit anti-PPARβ/δ (Abcam Inc. Cat. #8937, 1:500), and rabbit anti-PPARγ (Cell Signaling Technologies, Inc. Cat. #2443, 1:500) were used to measure protein expression of the PPAR isoforms.

    Techniques: Activity Assay, In Vivo, Isolation, Binding Assay, Enzyme-linked Immunosorbent Assay

    (A) Effects of S -allyl cysteine (SAC) administration on the gene expression and protein levels of Peroxisome proliferators-activated receptor (PPAR)-γ in the liver. Animals were treated as described in Fig. 1 . PPAR-γ mRNA expression was measured as described in Methods. Values are means ± SEM ( n = 5–6). # p

    Journal: Journal of Clinical Biochemistry and Nutrition

    Article Title: S-Allyl cysteine improves nonalcoholic fatty liver disease in type 2 diabetes Otsuka Long-Evans Tokushima Fatty rats via regulation of hepatic lipogenesis and glucose metabolism

    doi: 10.3164/jcbn.13-1

    Figure Lengend Snippet: (A) Effects of S -allyl cysteine (SAC) administration on the gene expression and protein levels of Peroxisome proliferators-activated receptor (PPAR)-γ in the liver. Animals were treated as described in Fig. 1 . PPAR-γ mRNA expression was measured as described in Methods. Values are means ± SEM ( n = 5–6). # p

    Article Snippet: Nuclear and cytosol proteins (15–20 µg) were analyzed by immunoblotting for PPAR-α and PPAR-γ (Cell Signaling Technology, Inc.) as described above.

    Techniques: Expressing