β actin antibody  (Boster Bio)


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    Name:
    Anti beta Actin ACTB Antibody
    Description:

    Catalog Number:
    A01263
    Price:
    399.0
    Category:
    Primary Antibodies
    Reactivity:
    Human
    Applications:
    IHC, WB
    Immunogen:
    beta-Actin Loading Control Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids 359-368 of Human beta Actin.
    Host:
    Rabbit
    Buy from Supplier


    Structured Review

    Boster Bio β actin antibody
    Anti beta Actin ACTB Antibody

    https://www.bioz.com/result/β actin antibody/product/Boster Bio
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    β actin antibody - by Bioz Stars, 2021-07
    94/100 stars

    Images

    1) Product Images from "Taurine attenuates oxidative stress and alleviates cardiac failure in type I diabetic rats"

    Article Title: Taurine attenuates oxidative stress and alleviates cardiac failure in type I diabetic rats

    Journal: Croatian Medical Journal

    doi: 10.3325/cmj.2013.54.171

    The levels of malondialdehyde and superoxide dismutase in cardiomyocyte from control and streptozotocin (STZ) diabetic rats treated with or without taurine (100 mg/kg). ( A ) malondialdehyde (MDA); ( B ) superoxide dismutase (SOD); ( C ) Representative gel blots of heme oxygenase-1 (HO-1) and β-actin (loading control) using specific antibodies; ( D ) HO-1 expression. Cont – control rats. Cont + taur – control rats with taurine. STZ – diabetic rats. STZ+taur – diabetic rats with taurine. The results are expressed as mean ± standard error of the mean. The number of animals per group was 8 for determination of MDA and HO-1 levels and SOD activity. * P
    Figure Legend Snippet: The levels of malondialdehyde and superoxide dismutase in cardiomyocyte from control and streptozotocin (STZ) diabetic rats treated with or without taurine (100 mg/kg). ( A ) malondialdehyde (MDA); ( B ) superoxide dismutase (SOD); ( C ) Representative gel blots of heme oxygenase-1 (HO-1) and β-actin (loading control) using specific antibodies; ( D ) HO-1 expression. Cont – control rats. Cont + taur – control rats with taurine. STZ – diabetic rats. STZ+taur – diabetic rats with taurine. The results are expressed as mean ± standard error of the mean. The number of animals per group was 8 for determination of MDA and HO-1 levels and SOD activity. * P

    Techniques Used: Multiple Displacement Amplification, Expressing, Activity Assay

    2) Product Images from "Acute Stress and Chronic Stress Change Brain-Derived Neurotrophic Factor (BDNF) and Tyrosine Kinase-Coupled Receptor (TrkB) Expression in Both Young and Aged Rat Hippocampus"

    Article Title: Acute Stress and Chronic Stress Change Brain-Derived Neurotrophic Factor (BDNF) and Tyrosine Kinase-Coupled Receptor (TrkB) Expression in Both Young and Aged Rat Hippocampus

    Journal: Yonsei Medical Journal

    doi: 10.3349/ymj.2010.51.5.661

    TrkB mRNA expression measured by semiquantitative RT-PCR in the control groups (unstressed, 0 min) and young and aged CMRS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for TrkB at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of TrkB mRNA in the young control and CMRS groups. (B) TrkB mRNA expression in the aged control and CMRS groups. (C) Line chart represented the results of quantitative analysis of TrkB mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. * p
    Figure Legend Snippet: TrkB mRNA expression measured by semiquantitative RT-PCR in the control groups (unstressed, 0 min) and young and aged CMRS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for TrkB at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of TrkB mRNA in the young control and CMRS groups. (B) TrkB mRNA expression in the aged control and CMRS groups. (C) Line chart represented the results of quantitative analysis of TrkB mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. * p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation

    BDNF mRNA expression detected by RT-PCR in the control groups (unstressed, 0 min) and young and aged CMRS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for BDNF at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of BDNF mRNA in the young control and CMRS groups. (B) BDNF mRNA expression in the aged control and CMRS groups. (C) Quantitative analysis of BDNF mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. * p
    Figure Legend Snippet: BDNF mRNA expression detected by RT-PCR in the control groups (unstressed, 0 min) and young and aged CMRS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for BDNF at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of BDNF mRNA in the young control and CMRS groups. (B) BDNF mRNA expression in the aged control and CMRS groups. (C) Quantitative analysis of BDNF mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. * p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation

    BDNF mRNA expression detected by semiquantitative RT-PCR in control groups (unstressed, 0 min) and young and aged AS groups after a period of different stress performance. Total RNA was isolated from hippocampus and assayed for BDNF at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms illustrating the expression of BDNF mRNA in the young control and AS groups. (B) Expression of BDNF mRNA in the aged AS group. (C) Quantitative analysis of BDNF mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of the β-actin (internal standard) band and expressed as the mean ± SEM. * p
    Figure Legend Snippet: BDNF mRNA expression detected by semiquantitative RT-PCR in control groups (unstressed, 0 min) and young and aged AS groups after a period of different stress performance. Total RNA was isolated from hippocampus and assayed for BDNF at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms illustrating the expression of BDNF mRNA in the young control and AS groups. (B) Expression of BDNF mRNA in the aged AS group. (C) Quantitative analysis of BDNF mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of the β-actin (internal standard) band and expressed as the mean ± SEM. * p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation

    TrkB mRNA expression measured by semiquantitative RT-PCR in the control groups (unstressed, 0 min) and young and aged AS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for TrkB mRNA at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of TrkB mRNA in the young control (0 min) and AS groups. (B) TrkB mRNA expression in the aged control and AS groups. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. n = 5-6 rats per each time point studied. TrkB, tyrosine kinase-coupled receptor; RT-PCR, reverse transcription-polymerase chain reaction; AS, acute stress.
    Figure Legend Snippet: TrkB mRNA expression measured by semiquantitative RT-PCR in the control groups (unstressed, 0 min) and young and aged AS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for TrkB mRNA at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of TrkB mRNA in the young control (0 min) and AS groups. (B) TrkB mRNA expression in the aged control and AS groups. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. n = 5-6 rats per each time point studied. TrkB, tyrosine kinase-coupled receptor; RT-PCR, reverse transcription-polymerase chain reaction; AS, acute stress.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation

    3) Product Images from "In vitro culture expansion impairs chondrogenic differentiation and the therapeutic effect of mesenchymal stem cells by regulating the unfolded protein response"

    Article Title: In vitro culture expansion impairs chondrogenic differentiation and the therapeutic effect of mesenchymal stem cells by regulating the unfolded protein response

    Journal: Journal of Biological Engineering

    doi: 10.1186/s13036-018-0119-2

    UPR-related gene/protein expression in four groups. a Quantitative RT-PCR for ATF6 , ATF4 , and XBP1 of the four groups in the cultured constructs on 0, 7, 14, and 21 days. These data were normalized to GAPDH . b Western blot results for ATF4, ATF6, XBP1, and COL2A1 at different culture times. COL2A1 was not detectable at 0 day; β-actin was used as a loading control. c Normalized expression of ATF4, ATF6, XBP1, and COL2A1 in response to Western blot analysis. d Immunofluorescence staining of ATF4, ATF6, and XBP1 at 0 days. Scale bar: 50 μm. The values were means ± S.D.; * indicate P
    Figure Legend Snippet: UPR-related gene/protein expression in four groups. a Quantitative RT-PCR for ATF6 , ATF4 , and XBP1 of the four groups in the cultured constructs on 0, 7, 14, and 21 days. These data were normalized to GAPDH . b Western blot results for ATF4, ATF6, XBP1, and COL2A1 at different culture times. COL2A1 was not detectable at 0 day; β-actin was used as a loading control. c Normalized expression of ATF4, ATF6, XBP1, and COL2A1 in response to Western blot analysis. d Immunofluorescence staining of ATF4, ATF6, and XBP1 at 0 days. Scale bar: 50 μm. The values were means ± S.D.; * indicate P

    Techniques Used: Expressing, Quantitative RT-PCR, Cell Culture, Construct, Western Blot, Immunofluorescence, Staining

    4) Product Images from "Pioglitazone ameliorates memory deficits in streptozotocin-induced diabetic mice by reducing brain β-amyloid through PPARγ activation"

    Article Title: Pioglitazone ameliorates memory deficits in streptozotocin-induced diabetic mice by reducing brain β-amyloid through PPARγ activation

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2013.11

    Pioglitazone suppresses RAGE and NF-κB p65 in the brain of STZ-induced diabetic mice. Representative immunoblots of RAGE, NF-κB p65, and β-actin (inner control) were obtained by Western blotting using the appropriate antibodies in the hippocampus and cerebral cortex (A). Quantification of RAGE or NF-κB p65 is expressed as a proportion (in percentage) relative to the control (B and C). Values shown are expressed as the mean±SEM. n =3. b P
    Figure Legend Snippet: Pioglitazone suppresses RAGE and NF-κB p65 in the brain of STZ-induced diabetic mice. Representative immunoblots of RAGE, NF-κB p65, and β-actin (inner control) were obtained by Western blotting using the appropriate antibodies in the hippocampus and cerebral cortex (A). Quantification of RAGE or NF-κB p65 is expressed as a proportion (in percentage) relative to the control (B and C). Values shown are expressed as the mean±SEM. n =3. b P

    Techniques Used: Mouse Assay, Western Blot

    Pioglitazone treatment prevents brain Aβ accumulation in STZ-induced diabetic mice. Aβ 1–40 (A) and Aβ 1–42 (B) levels in the hippocampus and cortex were assayed by ELISA. Representative immunoblots of Aβ 1–42 and β-actin (inner control) were obtained by Western blotting using the appropriate antibodies (C), and quantification of Aβ 1–42 is expressed as a proportion (in percentage) relative to the control (D). Values are expressed as the mean±SEM ( n =3–5). b P
    Figure Legend Snippet: Pioglitazone treatment prevents brain Aβ accumulation in STZ-induced diabetic mice. Aβ 1–40 (A) and Aβ 1–42 (B) levels in the hippocampus and cortex were assayed by ELISA. Representative immunoblots of Aβ 1–42 and β-actin (inner control) were obtained by Western blotting using the appropriate antibodies (C), and quantification of Aβ 1–42 is expressed as a proportion (in percentage) relative to the control (D). Values are expressed as the mean±SEM ( n =3–5). b P

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Western Blot

    Pioglitazone treatment reduces brain APP and BACE1 levels in STZ-induced diabetic mice. The expressions of APP and BACE1 in the hippocampus and cortex were detected by Western blotting. Representative immunoblots of APP, BACE1, and β-actin (inner control) are displayed (A), and quantification of APP or BACE1 is expressed as a proportion (in percentage) of the control (B and C). Values shown are expressed as the mean±SEM. n =3. b P
    Figure Legend Snippet: Pioglitazone treatment reduces brain APP and BACE1 levels in STZ-induced diabetic mice. The expressions of APP and BACE1 in the hippocampus and cortex were detected by Western blotting. Representative immunoblots of APP, BACE1, and β-actin (inner control) are displayed (A), and quantification of APP or BACE1 is expressed as a proportion (in percentage) of the control (B and C). Values shown are expressed as the mean±SEM. n =3. b P

    Techniques Used: Mouse Assay, Western Blot

    5) Product Images from "F10, a novel hydatidiform mole-associated gene, inhibits the paclitaxel sensitivity of A549 lung cancer cells by downregulating BAX and caspase-3"

    Article Title: F10, a novel hydatidiform mole-associated gene, inhibits the paclitaxel sensitivity of A549 lung cancer cells by downregulating BAX and caspase-3

    Journal: Oncology Letters

    doi: 10.3892/ol.2017.5749

    F10 reduced BAX and caspase-3 protein expression levels in paclitaxel-treated A549 cells. (A) The expression of BAX and caspase-3 was determined by a western blot analysis of paclitaxel-treated untransfected A549 cells, A549-mock cells or A549-F10 cells. The band signals of BAX and caspase-3 were normalized to that of β-actin. (B) The expression levels of BAX in paclitaxel-treated A549-F10 cells was lower compared with the untransfected A549 cells and A549-mock cells (*P
    Figure Legend Snippet: F10 reduced BAX and caspase-3 protein expression levels in paclitaxel-treated A549 cells. (A) The expression of BAX and caspase-3 was determined by a western blot analysis of paclitaxel-treated untransfected A549 cells, A549-mock cells or A549-F10 cells. The band signals of BAX and caspase-3 were normalized to that of β-actin. (B) The expression levels of BAX in paclitaxel-treated A549-F10 cells was lower compared with the untransfected A549 cells and A549-mock cells (*P

    Techniques Used: Expressing, Western Blot

    6) Product Images from "5,2′‐dibromo‐2,4′,5′‐trihydroxydiphenylmethanone attenuates LPS‐induced inflammation and ROS production in EA.hy926 cells via HMBOX1 induction, et al. 5,2′‐dibromo‐2,4′,5′‐trihydroxydiphenylmethanone attenuates LPS‐induced inflammation and ROS production in EA.hy926 cells via HMBOX1 induction"

    Article Title: 5,2′‐dibromo‐2,4′,5′‐trihydroxydiphenylmethanone attenuates LPS‐induced inflammation and ROS production in EA.hy926 cells via HMBOX1 induction, et al. 5,2′‐dibromo‐2,4′,5′‐trihydroxydiphenylmethanone attenuates LPS‐induced inflammation and ROS production in EA.hy926 cells via HMBOX1 induction

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.13948

    TDD‐mediated regulation of HMBOX1 expression in EA.hy926 cells. (A) HMBOX1 mRNA expression was analyzed by FQ‐PCR and normalized mRNA values relative to GAPDH levels were plotted. (B) Effect of TDD on HMBOX1 protein levels was evaluated by western blot with analysis of β‐actin expression as loading control. The data show the mean ± S.D. of three independent experiments. * P
    Figure Legend Snippet: TDD‐mediated regulation of HMBOX1 expression in EA.hy926 cells. (A) HMBOX1 mRNA expression was analyzed by FQ‐PCR and normalized mRNA values relative to GAPDH levels were plotted. (B) Effect of TDD on HMBOX1 protein levels was evaluated by western blot with analysis of β‐actin expression as loading control. The data show the mean ± S.D. of three independent experiments. * P

    Techniques Used: Expressing, Polymerase Chain Reaction, Western Blot

    7) Product Images from "Transcriptome sequencing of the naked mole rat (Heterocephalus glaber) and identification of hypoxia tolerance genes"

    Article Title: Transcriptome sequencing of the naked mole rat (Heterocephalus glaber) and identification of hypoxia tolerance genes

    Journal: Biology Open

    doi: 10.1242/bio.028548

    qRT-PCR and western blot verification of three selected DEGs. (A) qRT-PCR analysis of STMN1, JIP1 and JNK3 mRNA expression in muscle from NMRs under normoxic or hypoxic (5% O 2 ) condition. GAPDH was used as an internal reference. (B) STMN1, JIP1 and JNK3 protein expression was detected by western blot. β-Actin was used as an internal loading control. (C) Band density analysis of STMN1, JIP1 and JNK3 protein expression. The data represent means±s.e.m. of triplicate measurements. ★ P
    Figure Legend Snippet: qRT-PCR and western blot verification of three selected DEGs. (A) qRT-PCR analysis of STMN1, JIP1 and JNK3 mRNA expression in muscle from NMRs under normoxic or hypoxic (5% O 2 ) condition. GAPDH was used as an internal reference. (B) STMN1, JIP1 and JNK3 protein expression was detected by western blot. β-Actin was used as an internal loading control. (C) Band density analysis of STMN1, JIP1 and JNK3 protein expression. The data represent means±s.e.m. of triplicate measurements. ★ P

    Techniques Used: Quantitative RT-PCR, Western Blot, Expressing

    Expression of STMN1, JIP1 and JNK3 in muscle fibroblasts from NMRs before and after exposure to hypoxia. (A) Real-time PCR analysis of STMN1, JIP1 and JNK3 mRNA expression in muscle fibroblasts before and after exposure to hypoxia (5% O 2 ) for 12 h or 24 h. (B) Western blot detection of STMN1, JIP1 and JNK3 protein expression in fibroblasts before and after exposure to hypoxia (5% O 2 ) for 12 h or 24 h. β-actin was used as an internal loading control. (C,D) Band density analysis of STMN1, JIP1 and JNK3 protein expression in fibroblasts before and after exposure to hypoxia (5% O 2 ) for (C) 12 h or (D) 24 h. Data represent the mean±s.e.m. of five independent experiments. ★ P
    Figure Legend Snippet: Expression of STMN1, JIP1 and JNK3 in muscle fibroblasts from NMRs before and after exposure to hypoxia. (A) Real-time PCR analysis of STMN1, JIP1 and JNK3 mRNA expression in muscle fibroblasts before and after exposure to hypoxia (5% O 2 ) for 12 h or 24 h. (B) Western blot detection of STMN1, JIP1 and JNK3 protein expression in fibroblasts before and after exposure to hypoxia (5% O 2 ) for 12 h or 24 h. β-actin was used as an internal loading control. (C,D) Band density analysis of STMN1, JIP1 and JNK3 protein expression in fibroblasts before and after exposure to hypoxia (5% O 2 ) for (C) 12 h or (D) 24 h. Data represent the mean±s.e.m. of five independent experiments. ★ P

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

    8) Product Images from "Cisatracurium Retards Cell Migration and Invasion Upon Upregulation of p53 and Inhibits the Aggressiveness of Colorectal Cancer"

    Article Title: Cisatracurium Retards Cell Migration and Invasion Upon Upregulation of p53 and Inhibits the Aggressiveness of Colorectal Cancer

    Journal: Frontiers in Physiology

    doi: 10.3389/fphys.2018.00941

    (A–D) Cisatracurium alters migration and invasion regulatory genes transcription and protein expression levels in HCT116 cells. (A) Representative densities of SNAI-1, SLUG, E-Cadherin, CALD1, and β-actin proteins following western blot experiment. (B,C) Cluster bar charts of SNAI-1, SLUG, E-Cadherin, and CALD1 proteins expression and mRNA transcription levels of untreated and cisatracurium-treated (10 or 20 μM) HCT116 cells. β-Actin was used as internal control during western blot and qRT-PCR experiments. (D) Immunofluorescence was performed using FITC-labeled phalloidin, SNAI-1, SLUG, E-Cadherin, and CALD1 were stained with DAPI (Scale bar: 20 μm). Data are expressed as Mean ± SEM ( n = 3). ∗ p
    Figure Legend Snippet: (A–D) Cisatracurium alters migration and invasion regulatory genes transcription and protein expression levels in HCT116 cells. (A) Representative densities of SNAI-1, SLUG, E-Cadherin, CALD1, and β-actin proteins following western blot experiment. (B,C) Cluster bar charts of SNAI-1, SLUG, E-Cadherin, and CALD1 proteins expression and mRNA transcription levels of untreated and cisatracurium-treated (10 or 20 μM) HCT116 cells. β-Actin was used as internal control during western blot and qRT-PCR experiments. (D) Immunofluorescence was performed using FITC-labeled phalloidin, SNAI-1, SLUG, E-Cadherin, and CALD1 were stained with DAPI (Scale bar: 20 μm). Data are expressed as Mean ± SEM ( n = 3). ∗ p

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

    9) Product Images from "Antagonism of miR-21 Reverses Epithelial-Mesenchymal Transition and Cancer Stem Cell Phenotype through AKT/ERK1/2 Inactivation by Targeting PTEN"

    Article Title: Antagonism of miR-21 Reverses Epithelial-Mesenchymal Transition and Cancer Stem Cell Phenotype through AKT/ERK1/2 Inactivation by Targeting PTEN

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0039520

    Antagonism of miR-21 reversed EMT phenotype, as well as decreased cell migration and invasion. MDA-MB-231 cells were transfected with hsa-miR-21 antagomir or hsa-miR-21 antagomir control at a final concentration of 50 nmol for 48 h. (A) MDA-MB-231 cells were treated with hsa-miR-21 antagomir decreased the expression of miR-21, as compared to control groups (n1 = n2 = 3; p = 0.0015), by real-time RT-PCR analysis. (B-E) The mRNA levels of mesenchymal biomarkers (N-cadherin, Vimentin and alpha-SMA) and epithelial biomarker (E-cadherin) in MDA-MB-231/anti-miR-21 cells and MDA-MB-231/control cells, as measured by real-time RT-PCR analysis. The real-time RT-PCR reactions were performed in a 20 µl reaction volume in triplicate, simultaneously. (F, G) The relative protein levels of EMT markers in indicated cells were shown by Western blot analysis, and bands were semi-quantified using ImageJ software. Beta-actin was used as loading control. (H, I) The migratory and invasive properties of indicated cells were tested in migration and invasion assay in Transwell inserts. Penetrated cells were counted and analyzed in histogram. Data represent at least three experiments done in triplicate. (*indicates p
    Figure Legend Snippet: Antagonism of miR-21 reversed EMT phenotype, as well as decreased cell migration and invasion. MDA-MB-231 cells were transfected with hsa-miR-21 antagomir or hsa-miR-21 antagomir control at a final concentration of 50 nmol for 48 h. (A) MDA-MB-231 cells were treated with hsa-miR-21 antagomir decreased the expression of miR-21, as compared to control groups (n1 = n2 = 3; p = 0.0015), by real-time RT-PCR analysis. (B-E) The mRNA levels of mesenchymal biomarkers (N-cadherin, Vimentin and alpha-SMA) and epithelial biomarker (E-cadherin) in MDA-MB-231/anti-miR-21 cells and MDA-MB-231/control cells, as measured by real-time RT-PCR analysis. The real-time RT-PCR reactions were performed in a 20 µl reaction volume in triplicate, simultaneously. (F, G) The relative protein levels of EMT markers in indicated cells were shown by Western blot analysis, and bands were semi-quantified using ImageJ software. Beta-actin was used as loading control. (H, I) The migratory and invasive properties of indicated cells were tested in migration and invasion assay in Transwell inserts. Penetrated cells were counted and analyzed in histogram. Data represent at least three experiments done in triplicate. (*indicates p

    Techniques Used: Migration, Multiple Displacement Amplification, Transfection, Concentration Assay, Expressing, Quantitative RT-PCR, Biomarker Assay, Western Blot, Software, Invasion Assay

    Antagonism of miR-21 reversed CSC phenotype. (A, B) ALDH1 enzymatic activity (ALDH bright ) in established breast cancer MDA-MB-231/anti-miR-21 cells and MDA-MB-231/control cells (n1 = n2 = 3) were detected using the ALDEFLUOR assay. (C, D) The CD44 + /CD24 −/low phenotype in indicated cells (n1 = n2 = 3) were detected by FACS analysis. (E, F) The relative mRNA levels of ALDH1 and CD44 were detected by real time RT-PCR assay. (G, H) The relative protein levels of ALDH1 and CD44 were detected by Western blot analysis. Beta-actin was used as loading control. (I, J) The number of mammospheres from 1000 MDA-MB-231/anti-miR-21 cells or MDA-MB-231/control cells was counted under microscope. All the data represent at least three experiments done in triplicate. (*indicates p
    Figure Legend Snippet: Antagonism of miR-21 reversed CSC phenotype. (A, B) ALDH1 enzymatic activity (ALDH bright ) in established breast cancer MDA-MB-231/anti-miR-21 cells and MDA-MB-231/control cells (n1 = n2 = 3) were detected using the ALDEFLUOR assay. (C, D) The CD44 + /CD24 −/low phenotype in indicated cells (n1 = n2 = 3) were detected by FACS analysis. (E, F) The relative mRNA levels of ALDH1 and CD44 were detected by real time RT-PCR assay. (G, H) The relative protein levels of ALDH1 and CD44 were detected by Western blot analysis. Beta-actin was used as loading control. (I, J) The number of mammospheres from 1000 MDA-MB-231/anti-miR-21 cells or MDA-MB-231/control cells was counted under microscope. All the data represent at least three experiments done in triplicate. (*indicates p

    Techniques Used: Activity Assay, Multiple Displacement Amplification, FACS, Quantitative RT-PCR, Western Blot, Microscopy

    MiR-21 regulated EMT and CSC phenotype through mediating AKT and ERK1/2 activation. Established MDA-MB-231/anti-miR-21 cells were transfected with hsa-miR-21 mimics at a concentration of 40 nmol for 72 h. Then the cells were trypsinized, and treated with LY294002 (20 µmol/l) or U0126 (10 µmol/l) for 24 h. (A-E) The relative protein levels of p-AKT and AKT (A), p-ERK and ERK (A), EMT markers (B), CSC surface markers (C), as well as PTEN (D) from blank control, LY294002, and U0126 treatment groups were shown, and semi-quantified (E) as stated before. Beta-actin or GAPDH was used as loading control. (*indicates p
    Figure Legend Snippet: MiR-21 regulated EMT and CSC phenotype through mediating AKT and ERK1/2 activation. Established MDA-MB-231/anti-miR-21 cells were transfected with hsa-miR-21 mimics at a concentration of 40 nmol for 72 h. Then the cells were trypsinized, and treated with LY294002 (20 µmol/l) or U0126 (10 µmol/l) for 24 h. (A-E) The relative protein levels of p-AKT and AKT (A), p-ERK and ERK (A), EMT markers (B), CSC surface markers (C), as well as PTEN (D) from blank control, LY294002, and U0126 treatment groups were shown, and semi-quantified (E) as stated before. Beta-actin or GAPDH was used as loading control. (*indicates p

    Techniques Used: Activation Assay, Multiple Displacement Amplification, Transfection, Concentration Assay

    Hsa-miR-21 mimics induced EMT and CSC phenotype, accompanied with PTEN down-regulation and AKT/ERK1/2 activation. Established MDA-MB-231/anti-miR-21 cells were transfected with hsa-miR-21 mimics at a concentration of 40 nmol for 72 h. (A) MDA-MB-231/anti-miR-21 cells were treated with hsa-miR-21 mimics elevated the expression of miR-21, as compared to control groups (n1 = n2 = 3; p = 0.00373), by real-time RT-PCR analysis. (B-F) Protein levels of mesenchymal markers (N-cadherin, Vimentin and alpha-SMA) (B), epithelial marker (E-cadherin) (B), CSC markers (ALDH1 and CD44) (C), PTEN (D), p-AKT and AKT (E), as well as p-ERK1/2 and ERK1/2 (E) in indicated cells were measured by Western blot analysis, and bands were semi-quantified using ImageJ software (F). Beta-actin or GAPDH was used as loading control. (*indicates p
    Figure Legend Snippet: Hsa-miR-21 mimics induced EMT and CSC phenotype, accompanied with PTEN down-regulation and AKT/ERK1/2 activation. Established MDA-MB-231/anti-miR-21 cells were transfected with hsa-miR-21 mimics at a concentration of 40 nmol for 72 h. (A) MDA-MB-231/anti-miR-21 cells were treated with hsa-miR-21 mimics elevated the expression of miR-21, as compared to control groups (n1 = n2 = 3; p = 0.00373), by real-time RT-PCR analysis. (B-F) Protein levels of mesenchymal markers (N-cadherin, Vimentin and alpha-SMA) (B), epithelial marker (E-cadherin) (B), CSC markers (ALDH1 and CD44) (C), PTEN (D), p-AKT and AKT (E), as well as p-ERK1/2 and ERK1/2 (E) in indicated cells were measured by Western blot analysis, and bands were semi-quantified using ImageJ software (F). Beta-actin or GAPDH was used as loading control. (*indicates p

    Techniques Used: Activation Assay, Multiple Displacement Amplification, Transfection, Concentration Assay, Expressing, Quantitative RT-PCR, Marker, Western Blot, Software

    PTEN was the downstream target of miR-21 during reversing EMT and CSC phenotype . MDA-MB-231 cells were transfected with siPTEN or the scrambled control SsiPTEN at a final concentration of 50 nmol for 24 h. Then the cells were transfected with hsa-miR-21 antagomir or negative control at a final concentration of 50 nmol for 72 h. Cells were trypsinized, and the relative protein levels of PTEN (A), EMT markers (B), CSC surface markers (C), p-AKT and AKT (D), as well as p-ERK and ERK (D) were measured and semi-quantified (E) as stated before. The representative plugs from treatments of SsiPTEN plus miR-21 antagomir control, SsiPTEN plus miR-21 antagomir, and siPTEN plus miR-21 antagomir were shown in the picture. Beta-actin or GAPDH was used as loading control. (*indicates p
    Figure Legend Snippet: PTEN was the downstream target of miR-21 during reversing EMT and CSC phenotype . MDA-MB-231 cells were transfected with siPTEN or the scrambled control SsiPTEN at a final concentration of 50 nmol for 24 h. Then the cells were transfected with hsa-miR-21 antagomir or negative control at a final concentration of 50 nmol for 72 h. Cells were trypsinized, and the relative protein levels of PTEN (A), EMT markers (B), CSC surface markers (C), p-AKT and AKT (D), as well as p-ERK and ERK (D) were measured and semi-quantified (E) as stated before. The representative plugs from treatments of SsiPTEN plus miR-21 antagomir control, SsiPTEN plus miR-21 antagomir, and siPTEN plus miR-21 antagomir were shown in the picture. Beta-actin or GAPDH was used as loading control. (*indicates p

    Techniques Used: Multiple Displacement Amplification, Transfection, Concentration Assay, Negative Control

    10) Product Images from "Phosphatidylinositol 3-kinase affects mitochondrial function in part through inducing peroxisome proliferator-activated receptor ? coactivator-1? expression"

    Article Title: Phosphatidylinositol 3-kinase affects mitochondrial function in part through inducing peroxisome proliferator-activated receptor ? coactivator-1? expression

    Journal: British Journal of Pharmacology

    doi: 10.1111/j.1476-5381.2010.01105.x

    Suppressing PI3K activity selectively suppresses the expression of PGC-1β. (A) ERRs and PGC-1 s protein expression levels were assayed by Western blots with β-actin as an internal control. (B) PGC-1β gene expression levels were
    Figure Legend Snippet: Suppressing PI3K activity selectively suppresses the expression of PGC-1β. (A) ERRs and PGC-1 s protein expression levels were assayed by Western blots with β-actin as an internal control. (B) PGC-1β gene expression levels were

    Techniques Used: Activity Assay, Expressing, Pyrolysis Gas Chromatography, Western Blot

    11) Product Images from "Inhibitory effect of SLIT2 on granulosa cell proliferation mediated by the CDC42-PAKs-ERK1/2 MAPK pathway in the prehierarchical follicles of the chicken ovary"

    Article Title: Inhibitory effect of SLIT2 on granulosa cell proliferation mediated by the CDC42-PAKs-ERK1/2 MAPK pathway in the prehierarchical follicles of the chicken ovary

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-27601-z

    Effects of the SLIT2 knockdown on the expression of the ROBO1 and ROBO2 genes. The granulosa cells were transfected with the SLIT2 -specific siRNA, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) The expression of the SLIT2 gene in the GCs with or without the interference of the specific siRNA was determined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values of the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SLIT2 protein in the GCs with or without the siRNA interference were detected by western blotting. β-actin was used as a loading control. ( C ) The influence of the SLIT2 knockdown on the ROBO1 and ROBO2 mRNA abundance in the granulosa cells. ( D ) The effects of the SLIT2 knockdown on the protein levels of ROBO1 and ROBO2. All blots were cropped, and the gels were run under the same experimental conditions. For each group, the superscript symbol above the bar indicates that the difference was significant compared to the control group **P
    Figure Legend Snippet: Effects of the SLIT2 knockdown on the expression of the ROBO1 and ROBO2 genes. The granulosa cells were transfected with the SLIT2 -specific siRNA, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) The expression of the SLIT2 gene in the GCs with or without the interference of the specific siRNA was determined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values of the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SLIT2 protein in the GCs with or without the siRNA interference were detected by western blotting. β-actin was used as a loading control. ( C ) The influence of the SLIT2 knockdown on the ROBO1 and ROBO2 mRNA abundance in the granulosa cells. ( D ) The effects of the SLIT2 knockdown on the protein levels of ROBO1 and ROBO2. All blots were cropped, and the gels were run under the same experimental conditions. For each group, the superscript symbol above the bar indicates that the difference was significant compared to the control group **P

    Techniques Used: Expressing, Transfection, Negative Control, Quantitative RT-PCR, Western Blot

    Regulation of the phosphorylation of B-RAF, RAF1, ERK1/2 and GC proliferation through the kinase PAKs. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids and co-infected with or without PAK1 , PAK2 and PAK3 specific siRNA. ( A ) The phosphorylation levels of B-RAF and RAF1 proteins in the GCs with or without the specific siRNA interference were detected by western blotting and normalized for loading by comparison to the signal of β-actin. ( B ) The phosphorylation levels of the ERK1/2 proteins in the GCs with and without the specific siRNA interference were detected by western blotting and normalized for loading by comparison to the signal of β-actin. The signal intensity of the phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 10 per mean ± SEM). ( C ) Proliferation levels of the GCs with and without the specific siRNA interference were examined using an EdU Cell Proliferation Assay Kit. The statistical significance is indicated with different superscript characters (P
    Figure Legend Snippet: Regulation of the phosphorylation of B-RAF, RAF1, ERK1/2 and GC proliferation through the kinase PAKs. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids and co-infected with or without PAK1 , PAK2 and PAK3 specific siRNA. ( A ) The phosphorylation levels of B-RAF and RAF1 proteins in the GCs with or without the specific siRNA interference were detected by western blotting and normalized for loading by comparison to the signal of β-actin. ( B ) The phosphorylation levels of the ERK1/2 proteins in the GCs with and without the specific siRNA interference were detected by western blotting and normalized for loading by comparison to the signal of β-actin. The signal intensity of the phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 10 per mean ± SEM). ( C ) Proliferation levels of the GCs with and without the specific siRNA interference were examined using an EdU Cell Proliferation Assay Kit. The statistical significance is indicated with different superscript characters (P

    Techniques Used: Transfection, Infection, Western Blot, Proliferation Assay

    Knockdown of SLIT2 enhances the phosphorylation levels of PAKs, RAFs and ERK1/2. The granulosa cells were transfected with the SLIT2-specific siRNA, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) An in vitro phosphorylation assay was performed by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( B , C ) The blotting signal intensity was quantified densitometrically after phosphorimaging (shown in A ) and normalized for loading by comparison to the signal of β-actin. The signal intensity of the targeted proteins or phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 5 per mean ± SEM). Five independent experiments were carried out in triplicate. The results are representative of at least three independent experiments. The statistical significance is marked with different superscript symbols **P
    Figure Legend Snippet: Knockdown of SLIT2 enhances the phosphorylation levels of PAKs, RAFs and ERK1/2. The granulosa cells were transfected with the SLIT2-specific siRNA, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) An in vitro phosphorylation assay was performed by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( B , C ) The blotting signal intensity was quantified densitometrically after phosphorimaging (shown in A ) and normalized for loading by comparison to the signal of β-actin. The signal intensity of the targeted proteins or phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 5 per mean ± SEM). Five independent experiments were carried out in triplicate. The results are representative of at least three independent experiments. The statistical significance is marked with different superscript symbols **P

    Techniques Used: Transfection, Negative Control, In Vitro, Phosphorylation Assay, Western Blot

    Effects of silencing SLIT2 on SRGAP1 expression and the GTPase activity of CDC42 and RAC1 in GCs. The granulosa cells were transfected with specific siRNAs targeting the SLIT2 gene, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) The expression of the SRGAP1 gene before and after the GCs were transfected with the specific siRNA for 48 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SRGAP1 protein in the GCs with and without specific siRNA interference (RNAi) was detected by western blotting. β-actin was used as a loading control. ( C ) The immunoprecipitation (IP) with the PBD antibody was revealed by immunoblotting (IB) using monoclonal anti-CDC42 or anti-RAC1 antibodies (in the right-hand column). −, negative control; +, SLIT2 silencing group. All blots were cropped, and the gels were run under the same experimental conditions. ( D ) The expression levels of GTP-bound CDC42 and RAC1 under SLIT2 silencing were determined by western blotting. β-actin was used as a loading control. For each group, the different superscript above the bar indicates that the difference was significant (P
    Figure Legend Snippet: Effects of silencing SLIT2 on SRGAP1 expression and the GTPase activity of CDC42 and RAC1 in GCs. The granulosa cells were transfected with specific siRNAs targeting the SLIT2 gene, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) The expression of the SRGAP1 gene before and after the GCs were transfected with the specific siRNA for 48 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SRGAP1 protein in the GCs with and without specific siRNA interference (RNAi) was detected by western blotting. β-actin was used as a loading control. ( C ) The immunoprecipitation (IP) with the PBD antibody was revealed by immunoblotting (IB) using monoclonal anti-CDC42 or anti-RAC1 antibodies (in the right-hand column). −, negative control; +, SLIT2 silencing group. All blots were cropped, and the gels were run under the same experimental conditions. ( D ) The expression levels of GTP-bound CDC42 and RAC1 under SLIT2 silencing were determined by western blotting. β-actin was used as a loading control. For each group, the different superscript above the bar indicates that the difference was significant (P

    Techniques Used: Expressing, Activity Assay, Transfection, Negative Control, Quantitative RT-PCR, Western Blot, Immunoprecipitation

    SLIT2 overexpression-induced reduction in the phosphorylation levels of PAKs, RAFs and ERK1/2. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The immunoprecipitants were analyzed by western blotting for an in vitro phosphorylation assay. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( B , C ) The blotting signal intensity was quantified densitometrically after phosphorimaging (shown in A ) and normalized for loading by comparison to the signal of β-actin. The signal intensity of the targeted proteins or phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 5 per mean ± SEM). Five independent experiments were carried out in triplicate. The results are representative of at least three independent experiments. significance is marked with different superscript symbols **P
    Figure Legend Snippet: SLIT2 overexpression-induced reduction in the phosphorylation levels of PAKs, RAFs and ERK1/2. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The immunoprecipitants were analyzed by western blotting for an in vitro phosphorylation assay. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( B , C ) The blotting signal intensity was quantified densitometrically after phosphorimaging (shown in A ) and normalized for loading by comparison to the signal of β-actin. The signal intensity of the targeted proteins or phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 5 per mean ± SEM). Five independent experiments were carried out in triplicate. The results are representative of at least three independent experiments. significance is marked with different superscript symbols **P

    Techniques Used: Over Expression, Transfection, Plasmid Preparation, Negative Control, Western Blot, In Vitro, Phosphorylation Assay

    Effects of overexpressing SLIT2 on the expression of the ROBO1, ROBO2 , ROBO3 and ROBO4 genes. The granulosa cells were transfected with a reconstructed pYr-adshuttle-4-SLIT2 vector, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The expression of the SLIT2 gene before and after the GCs were transfected with the pYr-adshuttle-4-SLIT2 expression vector for 24 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SLIT2 protein in the GCs before and after the transfection with the pYr-adshuttle-4-SLIT2 vector were detected by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( C ) The influence of the SLIT2 overexpression on the ROBO1 , ROBO2, ROBO3 and ROBO4 mRNA abundance in the granulosa cells from the prehierarchical follicles (6 to 8 mm in diameter) was examined. ( D ) The effects of the SLIT2 overexpression on the protein levels of ROBO1 and ROBO2. For each group, the superscript symbol above the bar indicates that the difference was significant compared to the control group **P
    Figure Legend Snippet: Effects of overexpressing SLIT2 on the expression of the ROBO1, ROBO2 , ROBO3 and ROBO4 genes. The granulosa cells were transfected with a reconstructed pYr-adshuttle-4-SLIT2 vector, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The expression of the SLIT2 gene before and after the GCs were transfected with the pYr-adshuttle-4-SLIT2 expression vector for 24 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SLIT2 protein in the GCs before and after the transfection with the pYr-adshuttle-4-SLIT2 vector were detected by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( C ) The influence of the SLIT2 overexpression on the ROBO1 , ROBO2, ROBO3 and ROBO4 mRNA abundance in the granulosa cells from the prehierarchical follicles (6 to 8 mm in diameter) was examined. ( D ) The effects of the SLIT2 overexpression on the protein levels of ROBO1 and ROBO2. For each group, the superscript symbol above the bar indicates that the difference was significant compared to the control group **P

    Techniques Used: Expressing, Transfection, Plasmid Preparation, Negative Control, Quantitative RT-PCR, Western Blot, Over Expression

    Effects of the overexpressed SLIT2 on SRGAP1 expression and GTPase activity of CDC42 and RAC1 in GCs. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The expression of the SRGAP1 gene before and after the GCs were transfected with the pYr-adshuttle-4-SLIT2 expression vector for 24 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SRGAP1 protein in the GCs before and after the transfection with the pYr-adshuttle-4-SLIT2 vector was detected by western blotting. β-actin was used as a loading control. ( C ) The coimmunoprecipitation of CDC42 and RAC1 with the recombinant GST-PBD in vitro . The stimulation by the SLIT2 overexpression of GTP-bound CDC42 and RAC1 in the granulosa cells from the prehierarchical follicles (6 to 8 mm in diameter) was examined by western blotting after performing a GST pull-down assay in which cell lysates were incubated with glutathione S-transferase (GST) and the recombinant GST-PBD. The immunoprecipitation (IP) with the PBD antibody was revealed by immunoblotting (IB) using monoclonal anti-CDC42 or anti-RAC1 antibodies (in the right-hand column). −, negative control; +, SLIT2 overexpressed group. All blots were cropped, and the gels were run under the same experimental conditions. ( D ) The effects of the SLIT2 overexpression on the expression levels of GTP-bound CDC42 and RAC1 were detected by western blotting. β-actin was used as a loading control. For each group, the different superscript above the bar indicates that the difference was significant (P
    Figure Legend Snippet: Effects of the overexpressed SLIT2 on SRGAP1 expression and GTPase activity of CDC42 and RAC1 in GCs. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The expression of the SRGAP1 gene before and after the GCs were transfected with the pYr-adshuttle-4-SLIT2 expression vector for 24 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SRGAP1 protein in the GCs before and after the transfection with the pYr-adshuttle-4-SLIT2 vector was detected by western blotting. β-actin was used as a loading control. ( C ) The coimmunoprecipitation of CDC42 and RAC1 with the recombinant GST-PBD in vitro . The stimulation by the SLIT2 overexpression of GTP-bound CDC42 and RAC1 in the granulosa cells from the prehierarchical follicles (6 to 8 mm in diameter) was examined by western blotting after performing a GST pull-down assay in which cell lysates were incubated with glutathione S-transferase (GST) and the recombinant GST-PBD. The immunoprecipitation (IP) with the PBD antibody was revealed by immunoblotting (IB) using monoclonal anti-CDC42 or anti-RAC1 antibodies (in the right-hand column). −, negative control; +, SLIT2 overexpressed group. All blots were cropped, and the gels were run under the same experimental conditions. ( D ) The effects of the SLIT2 overexpression on the expression levels of GTP-bound CDC42 and RAC1 were detected by western blotting. β-actin was used as a loading control. For each group, the different superscript above the bar indicates that the difference was significant (P

    Techniques Used: Expressing, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Quantitative RT-PCR, Western Blot, Recombinant, In Vitro, Over Expression, Pull Down Assay, Incubation, Immunoprecipitation

    12) Product Images from "Nutlin-3-induced redistribution of chromatin-bound IFI16 in human hepatocellular carcinoma cells in vitro is associated with p53 activation"

    Article Title: Nutlin-3-induced redistribution of chromatin-bound IFI16 in human hepatocellular carcinoma cells in vitro is associated with p53 activation

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2014.106

    IFI16 subcellular localization regulated by Nutlin-3 in L02 cells. (A) IFI16 subcellular localization was detected in L02 cells using fluorescence microscopy (1000×). Cells were treated as described above and stained for IFI16 (green). Nuclei were counter-stained with DAPI (blue). Images were merged using Image-Pro plus 6.0. (B) Chromatin fractions were analyzed via Western blot in L02 cells. H2b and β-actin served as the quality control for the nuclear fraction and the cytoplasmic fraction, respectively. S1, cytoplasmic proteins; S2, soluble nuclear proteins; P2, chromatin-enriched sediment; WCE, whole-cell extracts.
    Figure Legend Snippet: IFI16 subcellular localization regulated by Nutlin-3 in L02 cells. (A) IFI16 subcellular localization was detected in L02 cells using fluorescence microscopy (1000×). Cells were treated as described above and stained for IFI16 (green). Nuclei were counter-stained with DAPI (blue). Images were merged using Image-Pro plus 6.0. (B) Chromatin fractions were analyzed via Western blot in L02 cells. H2b and β-actin served as the quality control for the nuclear fraction and the cytoplasmic fraction, respectively. S1, cytoplasmic proteins; S2, soluble nuclear proteins; P2, chromatin-enriched sediment; WCE, whole-cell extracts.

    Techniques Used: Fluorescence, Microscopy, Staining, Western Blot

    Nutlin-3 causes DNA DSB damage in SMMC-7721 cells. (A) Nutlin-3 increased γH2AX expression level. β-Actin served as the loading control. (B) The relative expression levels of γH2AX. The value represents the mean±SD derived from triplicate tests. b P
    Figure Legend Snippet: Nutlin-3 causes DNA DSB damage in SMMC-7721 cells. (A) Nutlin-3 increased γH2AX expression level. β-Actin served as the loading control. (B) The relative expression levels of γH2AX. The value represents the mean±SD derived from triplicate tests. b P

    Techniques Used: Expressing, Derivative Assay

    13) Product Images from "Nutlin-3-induced redistribution of chromatin-bound IFI16 in human hepatocellular carcinoma cells in vitro is associated with p53 activation"

    Article Title: Nutlin-3-induced redistribution of chromatin-bound IFI16 in human hepatocellular carcinoma cells in vitro is associated with p53 activation

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2014.106

    IFI16 subcellular localization regulated by Nutlin-3 in L02 cells. (A) IFI16 subcellular localization was detected in L02 cells using fluorescence microscopy (1000×). Cells were treated as described above and stained for IFI16 (green). Nuclei were counter-stained with DAPI (blue). Images were merged using Image-Pro plus 6.0. (B) Chromatin fractions were analyzed via Western blot in L02 cells. H2b and β-actin served as the quality control for the nuclear fraction and the cytoplasmic fraction, respectively. S1, cytoplasmic proteins; S2, soluble nuclear proteins; P2, chromatin-enriched sediment; WCE, whole-cell extracts.
    Figure Legend Snippet: IFI16 subcellular localization regulated by Nutlin-3 in L02 cells. (A) IFI16 subcellular localization was detected in L02 cells using fluorescence microscopy (1000×). Cells were treated as described above and stained for IFI16 (green). Nuclei were counter-stained with DAPI (blue). Images were merged using Image-Pro plus 6.0. (B) Chromatin fractions were analyzed via Western blot in L02 cells. H2b and β-actin served as the quality control for the nuclear fraction and the cytoplasmic fraction, respectively. S1, cytoplasmic proteins; S2, soluble nuclear proteins; P2, chromatin-enriched sediment; WCE, whole-cell extracts.

    Techniques Used: Fluorescence, Microscopy, Staining, Western Blot

    Nutlin-3 causes DNA DSB damage in SMMC-7721 cells. (A) Nutlin-3 increased γH2AX expression level. β-Actin served as the loading control. (B) The relative expression levels of γH2AX. The value represents the mean±SD derived from triplicate tests. b P
    Figure Legend Snippet: Nutlin-3 causes DNA DSB damage in SMMC-7721 cells. (A) Nutlin-3 increased γH2AX expression level. β-Actin served as the loading control. (B) The relative expression levels of γH2AX. The value represents the mean±SD derived from triplicate tests. b P

    Techniques Used: Expressing, Derivative Assay

    14) Product Images from "5,2′‐dibromo‐2,4′,5′‐trihydroxydiphenylmethanone attenuates LPS‐induced inflammation and ROS production in EA.hy926 cells via HMBOX1 induction, et al. 5,2′‐dibromo‐2,4′,5′‐trihydroxydiphenylmethanone attenuates LPS‐induced inflammation and ROS production in EA.hy926 cells via HMBOX1 induction"

    Article Title: 5,2′‐dibromo‐2,4′,5′‐trihydroxydiphenylmethanone attenuates LPS‐induced inflammation and ROS production in EA.hy926 cells via HMBOX1 induction, et al. 5,2′‐dibromo‐2,4′,5′‐trihydroxydiphenylmethanone attenuates LPS‐induced inflammation and ROS production in EA.hy926 cells via HMBOX1 induction

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.13948

    Effects of TDD on LPS ‐stimulated monocyte adhesion, adhesion molecule and pro‐inflammatory cytokines expression. (A) THP ‐1 cells were labeled with the fluorescent probe and the adhesion was determined. (B) The gene expression of VCAM ‐1, ICAM ‐1 and E‐selection levels were determined by real‐time PCR and normalized to GAPDH . (C) The expression of VCAM ‐1, ICAM ‐1 and E‐selection were measured by Western blot assay. The β‐actin protein level was considered as an internal control. (D) The production of TNF ‐α and IL ‐6 were measured by ELISA analysis. (E) The mRNA expression of IL ‐6, IL ‐1β, MCP ‐1 and TNF ‐α were determined by FQ ‐ PCR . (F, G) Effects of TDD on LPS ‐stimulated the IL ‐6, IL ‐1β, MCP ‐1, TNF ‐α, ICAM ‐1, VCAM ‐1 and E‐selection mRNA expression were determined by FQ ‐ PCR with or without transfecting HMBOX 1 si RNA . The data show the mean ± SD of three independent experiments. * P
    Figure Legend Snippet: Effects of TDD on LPS ‐stimulated monocyte adhesion, adhesion molecule and pro‐inflammatory cytokines expression. (A) THP ‐1 cells were labeled with the fluorescent probe and the adhesion was determined. (B) The gene expression of VCAM ‐1, ICAM ‐1 and E‐selection levels were determined by real‐time PCR and normalized to GAPDH . (C) The expression of VCAM ‐1, ICAM ‐1 and E‐selection were measured by Western blot assay. The β‐actin protein level was considered as an internal control. (D) The production of TNF ‐α and IL ‐6 were measured by ELISA analysis. (E) The mRNA expression of IL ‐6, IL ‐1β, MCP ‐1 and TNF ‐α were determined by FQ ‐ PCR . (F, G) Effects of TDD on LPS ‐stimulated the IL ‐6, IL ‐1β, MCP ‐1, TNF ‐α, ICAM ‐1, VCAM ‐1 and E‐selection mRNA expression were determined by FQ ‐ PCR with or without transfecting HMBOX 1 si RNA . The data show the mean ± SD of three independent experiments. * P

    Techniques Used: Expressing, Labeling, Selection, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction

    TDD‐mediated regulation of HMBOX1 expression in EA.hy926 cells. (A) HMBOX1 mRNA expression was analyzed by FQ‐PCR and normalized mRNA values relative to GAPDH levels were plotted. (B) Effect of TDD on HMBOX1 protein levels was evaluated by western blot with analysis of β‐actin expression as loading control. The data show the mean ± S.D. of three independent experiments. * P
    Figure Legend Snippet: TDD‐mediated regulation of HMBOX1 expression in EA.hy926 cells. (A) HMBOX1 mRNA expression was analyzed by FQ‐PCR and normalized mRNA values relative to GAPDH levels were plotted. (B) Effect of TDD on HMBOX1 protein levels was evaluated by western blot with analysis of β‐actin expression as loading control. The data show the mean ± S.D. of three independent experiments. * P

    Techniques Used: Expressing, Polymerase Chain Reaction, Western Blot

    Effects of TDD on nuclear translocation of NF ‐κB/p65 via HMBOX 1 activation in LPS ‐stimulated EA .hy926 Cells. NE , nuclear extracts; CE , cytoplasmic extracts. (A) The cells were treated with TDD for 4 h before addition of LPS (1 μg/ mL ) for another 2 h. Cytoplasmic and nuclear levels of NF ‐κB p65 were detected by Western blotting to analyze the translocation of NF ‐κB. Lamin B and β‐actin were used as loading controls for nuclear and cytosolic protein fractions, respectively. (B) Immunofluorescent imaging shows a TDD ‐mediated suppression of LPS ‐mediated nuclear translocation of NF ‐κB p65 in EA .hy926 Cells. The arrow indicates the position of NF ‐Κb p65 (magnification, ×1000). (C) Effects of TDD on nuclear translocation of NF ‐κB/p65 in LPS ‐stimulated EA .hy926 Cells with or without transfecting HMBOX 1 si RNA . (D) Effects of NF ‐κB, ERK 1/2 and p38 MAPK inhibitors on the transcription of IL ‐6, IL ‐1β, MCP ‐1 and TNF ‐α in LPS ‐stimulated EA .hy926 cells. The data show the mean ± SD of three independent experiments. * P
    Figure Legend Snippet: Effects of TDD on nuclear translocation of NF ‐κB/p65 via HMBOX 1 activation in LPS ‐stimulated EA .hy926 Cells. NE , nuclear extracts; CE , cytoplasmic extracts. (A) The cells were treated with TDD for 4 h before addition of LPS (1 μg/ mL ) for another 2 h. Cytoplasmic and nuclear levels of NF ‐κB p65 were detected by Western blotting to analyze the translocation of NF ‐κB. Lamin B and β‐actin were used as loading controls for nuclear and cytosolic protein fractions, respectively. (B) Immunofluorescent imaging shows a TDD ‐mediated suppression of LPS ‐mediated nuclear translocation of NF ‐κB p65 in EA .hy926 Cells. The arrow indicates the position of NF ‐Κb p65 (magnification, ×1000). (C) Effects of TDD on nuclear translocation of NF ‐κB/p65 in LPS ‐stimulated EA .hy926 Cells with or without transfecting HMBOX 1 si RNA . (D) Effects of NF ‐κB, ERK 1/2 and p38 MAPK inhibitors on the transcription of IL ‐6, IL ‐1β, MCP ‐1 and TNF ‐α in LPS ‐stimulated EA .hy926 cells. The data show the mean ± SD of three independent experiments. * P

    Techniques Used: Translocation Assay, Activation Assay, Western Blot, Imaging

    15) Product Images from "Azeliragon ameliorates Alzheimer's disease via the Janus tyrosine kinase and signal transducer and activator of transcription signaling pathway"

    Article Title: Azeliragon ameliorates Alzheimer's disease via the Janus tyrosine kinase and signal transducer and activator of transcription signaling pathway

    Journal: Clinics

    doi: 10.6061/clinics/2021/e2348

    (A) Western blotting assay for detecting the expression of NLRP1 and NeuN. (B, C) Quantification of the expression of NLRP1 and NeuN. (D) NLRP1/NeuN ratio. (E) Immunofluorescence assay for detecting the expression of NLRP1 and NeuN (×400) and (F) NLRP1/NeuN (+) cells in samples from the sham, Aβ1-42, and Aβ1-42+LPS groups. Protein levels were normalized to those of β-actin (Aβ1-42 vs . sham group, ** p
    Figure Legend Snippet: (A) Western blotting assay for detecting the expression of NLRP1 and NeuN. (B, C) Quantification of the expression of NLRP1 and NeuN. (D) NLRP1/NeuN ratio. (E) Immunofluorescence assay for detecting the expression of NLRP1 and NeuN (×400) and (F) NLRP1/NeuN (+) cells in samples from the sham, Aβ1-42, and Aβ1-42+LPS groups. Protein levels were normalized to those of β-actin (Aβ1-42 vs . sham group, ** p

    Techniques Used: Western Blot, Expressing, Immunofluorescence

    16) Product Images from "Identification of serum β-catenin as a biomarker in patients with HBV-related liver diseases"

    Article Title: Identification of serum β-catenin as a biomarker in patients with HBV-related liver diseases

    Journal: Journal of Translational Medicine

    doi: 10.1186/s12967-018-1645-x

    Increased β-catenin expression in HBV-infected hepatic cells. a Western blot analysis of β-catenin expression in hepatic L02 cells transfected with and without pcDNA3.1-HBV (1.3 and 1.1) or its control pcDNA3.1 for 48 h. β-actin served as a loading control. b Immunofluorescence staining for β-catenin in L02 cells transfected with and without pcDNA3.1-HBV (1.3 and 1.1) or its control pcDNA3.1 for 48 h. White scale bars = 50 µm. c ELISA assay for β-catenin expression in L02 cells transfected with and without pcDNA3.1-HBV (1.3 and 1.1) or its control pcDNA3.1 for 48 h. d LDH assay for LDH from L02 cells transfected with and without pcDNA3.1-HBV (1.3 and 1.1) or its control pcDNA3.1 for 48 h. e Flow cytometry analysis for apoptotic L02 cells transfected with and without pcDNA3.1-HBV (1.3 and 1.1) or its control pcDNA3.1 for 72 h. f Apoptosis index for L02 cells with different treatment. Ns no statistical significance
    Figure Legend Snippet: Increased β-catenin expression in HBV-infected hepatic cells. a Western blot analysis of β-catenin expression in hepatic L02 cells transfected with and without pcDNA3.1-HBV (1.3 and 1.1) or its control pcDNA3.1 for 48 h. β-actin served as a loading control. b Immunofluorescence staining for β-catenin in L02 cells transfected with and without pcDNA3.1-HBV (1.3 and 1.1) or its control pcDNA3.1 for 48 h. White scale bars = 50 µm. c ELISA assay for β-catenin expression in L02 cells transfected with and without pcDNA3.1-HBV (1.3 and 1.1) or its control pcDNA3.1 for 48 h. d LDH assay for LDH from L02 cells transfected with and without pcDNA3.1-HBV (1.3 and 1.1) or its control pcDNA3.1 for 48 h. e Flow cytometry analysis for apoptotic L02 cells transfected with and without pcDNA3.1-HBV (1.3 and 1.1) or its control pcDNA3.1 for 72 h. f Apoptosis index for L02 cells with different treatment. Ns no statistical significance

    Techniques Used: Expressing, Infection, Western Blot, Transfection, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay, Flow Cytometry, Cytometry

    17) Product Images from "Nutlin-3 downregulates p53 phosphorylation on serine392 and induces apoptosis in hepatocellular carcinoma cells"

    Article Title: Nutlin-3 downregulates p53 phosphorylation on serine392 and induces apoptosis in hepatocellular carcinoma cells

    Journal: BMB Reports

    doi: 10.5483/BMBRep.2014.47.4.146

    Nutlin-3 induced apoptosis in two HCC cell lines. (A) Representative Flow Cytometric graphs of Annexin V-FITC/PI double staining treated with or without nutlin-3 (10 μM) as indicated in method. (B) The apoptotic percentage was quantified via Flow Cytometry under Annexin V-FITC/PI staining. Data are shown by means ± S.D. (n = 3), *P < 0.05 vs. NC group. (C) Western blots were performed for protein expression of Bax, Bcl-2, and caspase-3. β-actin was used as the loading control.
    Figure Legend Snippet: Nutlin-3 induced apoptosis in two HCC cell lines. (A) Representative Flow Cytometric graphs of Annexin V-FITC/PI double staining treated with or without nutlin-3 (10 μM) as indicated in method. (B) The apoptotic percentage was quantified via Flow Cytometry under Annexin V-FITC/PI staining. Data are shown by means ± S.D. (n = 3), *P < 0.05 vs. NC group. (C) Western blots were performed for protein expression of Bax, Bcl-2, and caspase-3. β-actin was used as the loading control.

    Techniques Used: Flow Cytometry, Double Staining, Cytometry, Staining, Western Blot, Expressing

    Nutlin-3 down-regulates the protein expression levels of phospho-Ser 392 -p53. (A) The protein expression of phospho-Ser 392 -p53 and p53. The cells were treated as indicated. β-actin served as the loading control. (B) The subcellular localization of p53 was detected under immunofluorescence microscope (1,000×). The cells were treated as indicated and stained for p53 (red). Nuclei were counterstained with DAPI (blue). Images were merged using Image-Pro plus 6.0. (C) The subcellular localization of p53 was detected by western blot analysis. Anti-H2b and β-actin antibodies were used as a loading control of nuclear and cytoplasmic proteins, respectively. (cyto) cytoplasmic proteins; (nuc) nuclear proteins. (D) Representative sequencing data identified p53- Ser 392 mutant from PCR products of SMMC- 7721 cells treated with or without Nutlin-3.
    Figure Legend Snippet: Nutlin-3 down-regulates the protein expression levels of phospho-Ser 392 -p53. (A) The protein expression of phospho-Ser 392 -p53 and p53. The cells were treated as indicated. β-actin served as the loading control. (B) The subcellular localization of p53 was detected under immunofluorescence microscope (1,000×). The cells were treated as indicated and stained for p53 (red). Nuclei were counterstained with DAPI (blue). Images were merged using Image-Pro plus 6.0. (C) The subcellular localization of p53 was detected by western blot analysis. Anti-H2b and β-actin antibodies were used as a loading control of nuclear and cytoplasmic proteins, respectively. (cyto) cytoplasmic proteins; (nuc) nuclear proteins. (D) Representative sequencing data identified p53- Ser 392 mutant from PCR products of SMMC- 7721 cells treated with or without Nutlin-3.

    Techniques Used: Expressing, Immunofluorescence, Microscopy, Staining, Western Blot, Sequencing, Mutagenesis, Polymerase Chain Reaction

    18) Product Images from "Inhibitory effect of SLIT2 on granulosa cell proliferation mediated by the CDC42-PAKs-ERK1/2 MAPK pathway in the prehierarchical follicles of the chicken ovary"

    Article Title: Inhibitory effect of SLIT2 on granulosa cell proliferation mediated by the CDC42-PAKs-ERK1/2 MAPK pathway in the prehierarchical follicles of the chicken ovary

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-27601-z

    Effects of the SLIT2 knockdown on the expression of the ROBO1 and ROBO2 genes. The granulosa cells were transfected with the SLIT2 -specific siRNA, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) The expression of the SLIT2 gene in the GCs with or without the interference of the specific siRNA was determined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values of the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SLIT2 protein in the GCs with or without the siRNA interference were detected by western blotting. β-actin was used as a loading control. ( C ) The influence of the SLIT2 knockdown on the ROBO1 and ROBO2 mRNA abundance in the granulosa cells. ( D ) The effects of the SLIT2 knockdown on the protein levels of ROBO1 and ROBO2. All blots were cropped, and the gels were run under the same experimental conditions. For each group, the superscript symbol above the bar indicates that the difference was significant compared to the control group **P
    Figure Legend Snippet: Effects of the SLIT2 knockdown on the expression of the ROBO1 and ROBO2 genes. The granulosa cells were transfected with the SLIT2 -specific siRNA, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) The expression of the SLIT2 gene in the GCs with or without the interference of the specific siRNA was determined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values of the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SLIT2 protein in the GCs with or without the siRNA interference were detected by western blotting. β-actin was used as a loading control. ( C ) The influence of the SLIT2 knockdown on the ROBO1 and ROBO2 mRNA abundance in the granulosa cells. ( D ) The effects of the SLIT2 knockdown on the protein levels of ROBO1 and ROBO2. All blots were cropped, and the gels were run under the same experimental conditions. For each group, the superscript symbol above the bar indicates that the difference was significant compared to the control group **P

    Techniques Used: Expressing, Transfection, Negative Control, Quantitative RT-PCR, Western Blot

    Regulation of the phosphorylation of B-RAF, RAF1, ERK1/2 and GC proliferation through the kinase PAKs. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids and co-infected with or without PAK1 , PAK2 and PAK3 specific siRNA. ( A ) The phosphorylation levels of B-RAF and RAF1 proteins in the GCs with or without the specific siRNA interference were detected by western blotting and normalized for loading by comparison to the signal of β-actin. ( B ) The phosphorylation levels of the ERK1/2 proteins in the GCs with and without the specific siRNA interference were detected by western blotting and normalized for loading by comparison to the signal of β-actin. The signal intensity of the phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 10 per mean ± SEM). ( C ) Proliferation levels of the GCs with and without the specific siRNA interference were examined using an EdU Cell Proliferation Assay Kit. The statistical significance is indicated with different superscript characters (P
    Figure Legend Snippet: Regulation of the phosphorylation of B-RAF, RAF1, ERK1/2 and GC proliferation through the kinase PAKs. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids and co-infected with or without PAK1 , PAK2 and PAK3 specific siRNA. ( A ) The phosphorylation levels of B-RAF and RAF1 proteins in the GCs with or without the specific siRNA interference were detected by western blotting and normalized for loading by comparison to the signal of β-actin. ( B ) The phosphorylation levels of the ERK1/2 proteins in the GCs with and without the specific siRNA interference were detected by western blotting and normalized for loading by comparison to the signal of β-actin. The signal intensity of the phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 10 per mean ± SEM). ( C ) Proliferation levels of the GCs with and without the specific siRNA interference were examined using an EdU Cell Proliferation Assay Kit. The statistical significance is indicated with different superscript characters (P

    Techniques Used: Transfection, Infection, Western Blot, Proliferation Assay

    Effect of RAF RNAi on the SLIT2 overexpression-induced inhibition of the phosphorylation levels of MEK1/2 and ERK1/2. The granulosa cells were transfected with B-RAF and/or RAF1 specific siRNA, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) The expression of the B-RAF and RAF1 genes before and after the GCs were transfected with specific siRNA for 48 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10). ( B ) The immunoprecipitants were analyzed by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( C ) The protein expression levels of B-RAF and RAF1 in the GCs with and without specific siRNA interference were detected by western blotting and normalized for loading by comparison to the signal of β-actin. ( D ) The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids and co-infected with or without B-RAF and RAF1 specific siRNA. The negative controls refer to the phosphorylation levels of MEK1/2 and ERK1/2 shown in Fig. 8 . −, neither SLIT2 overexpression nor knockdown of RAFs ; +, SLIT2 overexpression or knockdown of RAFs . The signal intensity of the phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 5 per mean ± SEM). The statistical significance is indicated with different superscript characters (P
    Figure Legend Snippet: Effect of RAF RNAi on the SLIT2 overexpression-induced inhibition of the phosphorylation levels of MEK1/2 and ERK1/2. The granulosa cells were transfected with B-RAF and/or RAF1 specific siRNA, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) The expression of the B-RAF and RAF1 genes before and after the GCs were transfected with specific siRNA for 48 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10). ( B ) The immunoprecipitants were analyzed by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( C ) The protein expression levels of B-RAF and RAF1 in the GCs with and without specific siRNA interference were detected by western blotting and normalized for loading by comparison to the signal of β-actin. ( D ) The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids and co-infected with or without B-RAF and RAF1 specific siRNA. The negative controls refer to the phosphorylation levels of MEK1/2 and ERK1/2 shown in Fig. 8 . −, neither SLIT2 overexpression nor knockdown of RAFs ; +, SLIT2 overexpression or knockdown of RAFs . The signal intensity of the phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 5 per mean ± SEM). The statistical significance is indicated with different superscript characters (P

    Techniques Used: Over Expression, Inhibition, Transfection, Negative Control, Expressing, Quantitative RT-PCR, Western Blot, Infection

    Knockdown of SLIT2 enhances the phosphorylation levels of PAKs, RAFs and ERK1/2. The granulosa cells were transfected with the SLIT2-specific siRNA, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) An in vitro phosphorylation assay was performed by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( B , C ) The blotting signal intensity was quantified densitometrically after phosphorimaging (shown in A ) and normalized for loading by comparison to the signal of β-actin. The signal intensity of the targeted proteins or phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 5 per mean ± SEM). Five independent experiments were carried out in triplicate. The results are representative of at least three independent experiments. The statistical significance is marked with different superscript symbols **P
    Figure Legend Snippet: Knockdown of SLIT2 enhances the phosphorylation levels of PAKs, RAFs and ERK1/2. The granulosa cells were transfected with the SLIT2-specific siRNA, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) An in vitro phosphorylation assay was performed by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( B , C ) The blotting signal intensity was quantified densitometrically after phosphorimaging (shown in A ) and normalized for loading by comparison to the signal of β-actin. The signal intensity of the targeted proteins or phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 5 per mean ± SEM). Five independent experiments were carried out in triplicate. The results are representative of at least three independent experiments. The statistical significance is marked with different superscript symbols **P

    Techniques Used: Transfection, Negative Control, In Vitro, Phosphorylation Assay, Western Blot

    Effects of silencing SLIT2 on SRGAP1 expression and the GTPase activity of CDC42 and RAC1 in GCs. The granulosa cells were transfected with specific siRNAs targeting the SLIT2 gene, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) The expression of the SRGAP1 gene before and after the GCs were transfected with the specific siRNA for 48 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SRGAP1 protein in the GCs with and without specific siRNA interference (RNAi) was detected by western blotting. β-actin was used as a loading control. ( C ) The immunoprecipitation (IP) with the PBD antibody was revealed by immunoblotting (IB) using monoclonal anti-CDC42 or anti-RAC1 antibodies (in the right-hand column). −, negative control; +, SLIT2 silencing group. All blots were cropped, and the gels were run under the same experimental conditions. ( D ) The expression levels of GTP-bound CDC42 and RAC1 under SLIT2 silencing were determined by western blotting. β-actin was used as a loading control. For each group, the different superscript above the bar indicates that the difference was significant (P
    Figure Legend Snippet: Effects of silencing SLIT2 on SRGAP1 expression and the GTPase activity of CDC42 and RAC1 in GCs. The granulosa cells were transfected with specific siRNAs targeting the SLIT2 gene, scrambled siRNA (negative control) or no siRNA (blank control). ( A ) The expression of the SRGAP1 gene before and after the GCs were transfected with the specific siRNA for 48 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SRGAP1 protein in the GCs with and without specific siRNA interference (RNAi) was detected by western blotting. β-actin was used as a loading control. ( C ) The immunoprecipitation (IP) with the PBD antibody was revealed by immunoblotting (IB) using monoclonal anti-CDC42 or anti-RAC1 antibodies (in the right-hand column). −, negative control; +, SLIT2 silencing group. All blots were cropped, and the gels were run under the same experimental conditions. ( D ) The expression levels of GTP-bound CDC42 and RAC1 under SLIT2 silencing were determined by western blotting. β-actin was used as a loading control. For each group, the different superscript above the bar indicates that the difference was significant (P

    Techniques Used: Expressing, Activity Assay, Transfection, Negative Control, Quantitative RT-PCR, Western Blot, Immunoprecipitation

    SLIT2 overexpression-induced reduction in the phosphorylation levels of PAKs, RAFs and ERK1/2. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The immunoprecipitants were analyzed by western blotting for an in vitro phosphorylation assay. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( B , C ) The blotting signal intensity was quantified densitometrically after phosphorimaging (shown in A ) and normalized for loading by comparison to the signal of β-actin. The signal intensity of the targeted proteins or phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 5 per mean ± SEM). Five independent experiments were carried out in triplicate. The results are representative of at least three independent experiments. significance is marked with different superscript symbols **P
    Figure Legend Snippet: SLIT2 overexpression-induced reduction in the phosphorylation levels of PAKs, RAFs and ERK1/2. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The immunoprecipitants were analyzed by western blotting for an in vitro phosphorylation assay. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( B , C ) The blotting signal intensity was quantified densitometrically after phosphorimaging (shown in A ) and normalized for loading by comparison to the signal of β-actin. The signal intensity of the targeted proteins or phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 5 per mean ± SEM). Five independent experiments were carried out in triplicate. The results are representative of at least three independent experiments. significance is marked with different superscript symbols **P

    Techniques Used: Over Expression, Transfection, Plasmid Preparation, Negative Control, Western Blot, In Vitro, Phosphorylation Assay

    Effects of overexpressing SLIT2 on the expression of the ROBO1, ROBO2 , ROBO3 and ROBO4 genes. The granulosa cells were transfected with a reconstructed pYr-adshuttle-4-SLIT2 vector, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The expression of the SLIT2 gene before and after the GCs were transfected with the pYr-adshuttle-4-SLIT2 expression vector for 24 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SLIT2 protein in the GCs before and after the transfection with the pYr-adshuttle-4-SLIT2 vector were detected by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( C ) The influence of the SLIT2 overexpression on the ROBO1 , ROBO2, ROBO3 and ROBO4 mRNA abundance in the granulosa cells from the prehierarchical follicles (6 to 8 mm in diameter) was examined. ( D ) The effects of the SLIT2 overexpression on the protein levels of ROBO1 and ROBO2. For each group, the superscript symbol above the bar indicates that the difference was significant compared to the control group **P
    Figure Legend Snippet: Effects of overexpressing SLIT2 on the expression of the ROBO1, ROBO2 , ROBO3 and ROBO4 genes. The granulosa cells were transfected with a reconstructed pYr-adshuttle-4-SLIT2 vector, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The expression of the SLIT2 gene before and after the GCs were transfected with the pYr-adshuttle-4-SLIT2 expression vector for 24 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SLIT2 protein in the GCs before and after the transfection with the pYr-adshuttle-4-SLIT2 vector were detected by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. ( C ) The influence of the SLIT2 overexpression on the ROBO1 , ROBO2, ROBO3 and ROBO4 mRNA abundance in the granulosa cells from the prehierarchical follicles (6 to 8 mm in diameter) was examined. ( D ) The effects of the SLIT2 overexpression on the protein levels of ROBO1 and ROBO2. For each group, the superscript symbol above the bar indicates that the difference was significant compared to the control group **P

    Techniques Used: Expressing, Transfection, Plasmid Preparation, Negative Control, Quantitative RT-PCR, Western Blot, Over Expression

    Effects of the overexpressed SLIT2 on SRGAP1 expression and GTPase activity of CDC42 and RAC1 in GCs. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The expression of the SRGAP1 gene before and after the GCs were transfected with the pYr-adshuttle-4-SLIT2 expression vector for 24 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SRGAP1 protein in the GCs before and after the transfection with the pYr-adshuttle-4-SLIT2 vector was detected by western blotting. β-actin was used as a loading control. ( C ) The coimmunoprecipitation of CDC42 and RAC1 with the recombinant GST-PBD in vitro . The stimulation by the SLIT2 overexpression of GTP-bound CDC42 and RAC1 in the granulosa cells from the prehierarchical follicles (6 to 8 mm in diameter) was examined by western blotting after performing a GST pull-down assay in which cell lysates were incubated with glutathione S-transferase (GST) and the recombinant GST-PBD. The immunoprecipitation (IP) with the PBD antibody was revealed by immunoblotting (IB) using monoclonal anti-CDC42 or anti-RAC1 antibodies (in the right-hand column). −, negative control; +, SLIT2 overexpressed group. All blots were cropped, and the gels were run under the same experimental conditions. ( D ) The effects of the SLIT2 overexpression on the expression levels of GTP-bound CDC42 and RAC1 were detected by western blotting. β-actin was used as a loading control. For each group, the different superscript above the bar indicates that the difference was significant (P
    Figure Legend Snippet: Effects of the overexpressed SLIT2 on SRGAP1 expression and GTPase activity of CDC42 and RAC1 in GCs. The granulosa cells were transfected with the reconstructed pYr-adshuttle-4-SLIT2 plasmids, a pYr-adshuttle-4 empty vector (negative control) or no plasmid (blank control). ( A ) The expression of the SRGAP1 gene before and after the GCs were transfected with the pYr-adshuttle-4-SLIT2 expression vector for 24 h was examined by qRT-PCR. The mRNA expression was normalized to that of the 18S rRNA gene; the values on the bar graphs represent the mean ± SEM of 10 hens (n = 10) from a representative experiment. ( B ) The expression levels of the SRGAP1 protein in the GCs before and after the transfection with the pYr-adshuttle-4-SLIT2 vector was detected by western blotting. β-actin was used as a loading control. ( C ) The coimmunoprecipitation of CDC42 and RAC1 with the recombinant GST-PBD in vitro . The stimulation by the SLIT2 overexpression of GTP-bound CDC42 and RAC1 in the granulosa cells from the prehierarchical follicles (6 to 8 mm in diameter) was examined by western blotting after performing a GST pull-down assay in which cell lysates were incubated with glutathione S-transferase (GST) and the recombinant GST-PBD. The immunoprecipitation (IP) with the PBD antibody was revealed by immunoblotting (IB) using monoclonal anti-CDC42 or anti-RAC1 antibodies (in the right-hand column). −, negative control; +, SLIT2 overexpressed group. All blots were cropped, and the gels were run under the same experimental conditions. ( D ) The effects of the SLIT2 overexpression on the expression levels of GTP-bound CDC42 and RAC1 were detected by western blotting. β-actin was used as a loading control. For each group, the different superscript above the bar indicates that the difference was significant (P

    Techniques Used: Expressing, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Quantitative RT-PCR, Western Blot, Recombinant, In Vitro, Over Expression, Pull Down Assay, Incubation, Immunoprecipitation

    19) Product Images from "Effects of GGT and C-S Lyase on the Generation of Endogenous Formaldehyde in Lentinula edodes at Different Growth Stages"

    Article Title: Effects of GGT and C-S Lyase on the Generation of Endogenous Formaldehyde in Lentinula edodes at Different Growth Stages

    Journal: Molecules

    doi: 10.3390/molecules24234203

    ( A ) Protein levels of Leggt at five stages of growth. ( B ) Protein levels of Lecsl at five stages of growth. ( C ) Relative expression of Leggt and Lecsl during five growth stages. β-actin protein was used as loading control, and the expressions during the M stage were taken as 100%. M, mycelia; G, grey; YFB, young fruiting body; IFB, immature fruiting body; MFB, mature fruiting body. Error bars indicate standard deviation for three independent experiments. * p
    Figure Legend Snippet: ( A ) Protein levels of Leggt at five stages of growth. ( B ) Protein levels of Lecsl at five stages of growth. ( C ) Relative expression of Leggt and Lecsl during five growth stages. β-actin protein was used as loading control, and the expressions during the M stage were taken as 100%. M, mycelia; G, grey; YFB, young fruiting body; IFB, immature fruiting body; MFB, mature fruiting body. Error bars indicate standard deviation for three independent experiments. * p

    Techniques Used: Expressing, Standard Deviation

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    Article Snippet: The proteins were separated in sodium dodecyl sulfate (SDS-PAGE) and electroblotted onto PVDF membranes. .. The membranes were incubated overnight at 4 °C with appropriate primary antibodies against β-Actin (Boster, Wuhan, China), COL2A1 (Boster), XBP1 (Sigma, USA), ATF6 (Sigma) or ATF4 (Bioss, Beijing, China) after incubation in 1% BSA blocking buffer (Boster) for 1 h at room temperature. .. Immunoreactive signals were detected by secondary antibodies (Invitrogen, Carlsbad, CA, USA) using the Odyssey Infrared Imaging System (LI-COR).

    Blocking Assay:

    Article Title: In vitro culture expansion impairs chondrogenic differentiation and the therapeutic effect of mesenchymal stem cells by regulating the unfolded protein response
    Article Snippet: The proteins were separated in sodium dodecyl sulfate (SDS-PAGE) and electroblotted onto PVDF membranes. .. The membranes were incubated overnight at 4 °C with appropriate primary antibodies against β-Actin (Boster, Wuhan, China), COL2A1 (Boster), XBP1 (Sigma, USA), ATF6 (Sigma) or ATF4 (Bioss, Beijing, China) after incubation in 1% BSA blocking buffer (Boster) for 1 h at room temperature. .. Immunoreactive signals were detected by secondary antibodies (Invitrogen, Carlsbad, CA, USA) using the Odyssey Infrared Imaging System (LI-COR).

    Stripping Membranes:

    Article Title: Transcriptome sequencing of the naked mole rat (Heterocephalus glaber) and identification of hypoxia tolerance genes
    Article Snippet: Subsequently, the membranes were washed three times with Tris-buffered saline with Tween 20 (TBST) (5 min per wash) and incubated with a goat anti-rabbit IgG antibodies diluted 1:4000 in blotting buffer (5% skimned milk). .. Blots were stripped with stripping buffer and reprobed with anti-β-actin antibody (Wuhan Boster Biotech, Wuhan, China) at a dilution of 1:2000 to confirm equal protein loading among the samples. .. Finally, immunoreactive bands were visualized using the Kodak Gel Logic 4000 R Imaging System (Carestream, USA).

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    Boster Bio β actin antibody
    The levels of malondialdehyde and superoxide dismutase in cardiomyocyte from control and streptozotocin (STZ) diabetic rats treated with or without taurine (100 mg/kg). ( A ) malondialdehyde (MDA); ( B ) superoxide dismutase (SOD); ( C ) Representative gel blots of heme oxygenase-1 (HO-1) and <t>β-actin</t> (loading control) using specific antibodies; ( D ) HO-1 expression. Cont – control rats. Cont + taur – control rats with taurine. STZ – diabetic rats. STZ+taur – diabetic rats with taurine. The results are expressed as mean ± standard error of the mean. The number of animals per group was 8 for determination of MDA and HO-1 levels and SOD activity. * P
    β Actin Antibody, supplied by Boster Bio, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The levels of malondialdehyde and superoxide dismutase in cardiomyocyte from control and streptozotocin (STZ) diabetic rats treated with or without taurine (100 mg/kg). ( A ) malondialdehyde (MDA); ( B ) superoxide dismutase (SOD); ( C ) Representative gel blots of heme oxygenase-1 (HO-1) and β-actin (loading control) using specific antibodies; ( D ) HO-1 expression. Cont – control rats. Cont + taur – control rats with taurine. STZ – diabetic rats. STZ+taur – diabetic rats with taurine. The results are expressed as mean ± standard error of the mean. The number of animals per group was 8 for determination of MDA and HO-1 levels and SOD activity. * P

    Journal: Croatian Medical Journal

    Article Title: Taurine attenuates oxidative stress and alleviates cardiac failure in type I diabetic rats

    doi: 10.3325/cmj.2013.54.171

    Figure Lengend Snippet: The levels of malondialdehyde and superoxide dismutase in cardiomyocyte from control and streptozotocin (STZ) diabetic rats treated with or without taurine (100 mg/kg). ( A ) malondialdehyde (MDA); ( B ) superoxide dismutase (SOD); ( C ) Representative gel blots of heme oxygenase-1 (HO-1) and β-actin (loading control) using specific antibodies; ( D ) HO-1 expression. Cont – control rats. Cont + taur – control rats with taurine. STZ – diabetic rats. STZ+taur – diabetic rats with taurine. The results are expressed as mean ± standard error of the mean. The number of animals per group was 8 for determination of MDA and HO-1 levels and SOD activity. * P

    Article Snippet: Nitrocellulose blots were blocked by incubation in TBS-T (10 mM Tris-HCl, pH 7.5, 150 mM NaCl, and 0.1% Tween 20) containing 5% non-fat milk for 1 hour at room temperature and incubated with a rabbit polyclonal anti-HO-1, AKT/PKB, phospho-AKT/PKB, CTGF, β-actin antibody (1:500 dilution; Wuhan Boster Biotechnologies, Wuhan, China) overnight at 4°C.

    Techniques: Multiple Displacement Amplification, Expressing, Activity Assay

    TrkB mRNA expression measured by semiquantitative RT-PCR in the control groups (unstressed, 0 min) and young and aged CMRS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for TrkB at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of TrkB mRNA in the young control and CMRS groups. (B) TrkB mRNA expression in the aged control and CMRS groups. (C) Line chart represented the results of quantitative analysis of TrkB mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. * p

    Journal: Yonsei Medical Journal

    Article Title: Acute Stress and Chronic Stress Change Brain-Derived Neurotrophic Factor (BDNF) and Tyrosine Kinase-Coupled Receptor (TrkB) Expression in Both Young and Aged Rat Hippocampus

    doi: 10.3349/ymj.2010.51.5.661

    Figure Lengend Snippet: TrkB mRNA expression measured by semiquantitative RT-PCR in the control groups (unstressed, 0 min) and young and aged CMRS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for TrkB at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of TrkB mRNA in the young control and CMRS groups. (B) TrkB mRNA expression in the aged control and CMRS groups. (C) Line chart represented the results of quantitative analysis of TrkB mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. * p

    Article Snippet: Anti-BDNF antibody, anti-β-actin antibody, and the secondary antibody were purchased from Wuhan Boster Company ( http://www.boster.com.cn/ ).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation

    BDNF mRNA expression detected by RT-PCR in the control groups (unstressed, 0 min) and young and aged CMRS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for BDNF at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of BDNF mRNA in the young control and CMRS groups. (B) BDNF mRNA expression in the aged control and CMRS groups. (C) Quantitative analysis of BDNF mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. * p

    Journal: Yonsei Medical Journal

    Article Title: Acute Stress and Chronic Stress Change Brain-Derived Neurotrophic Factor (BDNF) and Tyrosine Kinase-Coupled Receptor (TrkB) Expression in Both Young and Aged Rat Hippocampus

    doi: 10.3349/ymj.2010.51.5.661

    Figure Lengend Snippet: BDNF mRNA expression detected by RT-PCR in the control groups (unstressed, 0 min) and young and aged CMRS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for BDNF at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of BDNF mRNA in the young control and CMRS groups. (B) BDNF mRNA expression in the aged control and CMRS groups. (C) Quantitative analysis of BDNF mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. * p

    Article Snippet: Anti-BDNF antibody, anti-β-actin antibody, and the secondary antibody were purchased from Wuhan Boster Company ( http://www.boster.com.cn/ ).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation

    BDNF mRNA expression detected by semiquantitative RT-PCR in control groups (unstressed, 0 min) and young and aged AS groups after a period of different stress performance. Total RNA was isolated from hippocampus and assayed for BDNF at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms illustrating the expression of BDNF mRNA in the young control and AS groups. (B) Expression of BDNF mRNA in the aged AS group. (C) Quantitative analysis of BDNF mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of the β-actin (internal standard) band and expressed as the mean ± SEM. * p

    Journal: Yonsei Medical Journal

    Article Title: Acute Stress and Chronic Stress Change Brain-Derived Neurotrophic Factor (BDNF) and Tyrosine Kinase-Coupled Receptor (TrkB) Expression in Both Young and Aged Rat Hippocampus

    doi: 10.3349/ymj.2010.51.5.661

    Figure Lengend Snippet: BDNF mRNA expression detected by semiquantitative RT-PCR in control groups (unstressed, 0 min) and young and aged AS groups after a period of different stress performance. Total RNA was isolated from hippocampus and assayed for BDNF at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms illustrating the expression of BDNF mRNA in the young control and AS groups. (B) Expression of BDNF mRNA in the aged AS group. (C) Quantitative analysis of BDNF mRNA at different time points after stress. The results were calculated as the intensity of the lane of each transcript over the intensity of the β-actin (internal standard) band and expressed as the mean ± SEM. * p

    Article Snippet: Anti-BDNF antibody, anti-β-actin antibody, and the secondary antibody were purchased from Wuhan Boster Company ( http://www.boster.com.cn/ ).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation

    TrkB mRNA expression measured by semiquantitative RT-PCR in the control groups (unstressed, 0 min) and young and aged AS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for TrkB mRNA at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of TrkB mRNA in the young control (0 min) and AS groups. (B) TrkB mRNA expression in the aged control and AS groups. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. n = 5-6 rats per each time point studied. TrkB, tyrosine kinase-coupled receptor; RT-PCR, reverse transcription-polymerase chain reaction; AS, acute stress.

    Journal: Yonsei Medical Journal

    Article Title: Acute Stress and Chronic Stress Change Brain-Derived Neurotrophic Factor (BDNF) and Tyrosine Kinase-Coupled Receptor (TrkB) Expression in Both Young and Aged Rat Hippocampus

    doi: 10.3349/ymj.2010.51.5.661

    Figure Lengend Snippet: TrkB mRNA expression measured by semiquantitative RT-PCR in the control groups (unstressed, 0 min) and young and aged AS groups after a period of different stress performance. Total RNA was isolated from the hippocampus and assayed for TrkB mRNA at 15, 30, 60, 180, 720 min after stress. (A) Representative electrophoretograms showing the expression of TrkB mRNA in the young control (0 min) and AS groups. (B) TrkB mRNA expression in the aged control and AS groups. The results were calculated as the intensity of the lane of each transcript over the intensity of β-actin (internal standard) band and expressed as the mean ± SEM. n = 5-6 rats per each time point studied. TrkB, tyrosine kinase-coupled receptor; RT-PCR, reverse transcription-polymerase chain reaction; AS, acute stress.

    Article Snippet: Anti-BDNF antibody, anti-β-actin antibody, and the secondary antibody were purchased from Wuhan Boster Company ( http://www.boster.com.cn/ ).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation

    UPR-related gene/protein expression in four groups. a Quantitative RT-PCR for ATF6 , ATF4 , and XBP1 of the four groups in the cultured constructs on 0, 7, 14, and 21 days. These data were normalized to GAPDH . b Western blot results for ATF4, ATF6, XBP1, and COL2A1 at different culture times. COL2A1 was not detectable at 0 day; β-actin was used as a loading control. c Normalized expression of ATF4, ATF6, XBP1, and COL2A1 in response to Western blot analysis. d Immunofluorescence staining of ATF4, ATF6, and XBP1 at 0 days. Scale bar: 50 μm. The values were means ± S.D.; * indicate P

    Journal: Journal of Biological Engineering

    Article Title: In vitro culture expansion impairs chondrogenic differentiation and the therapeutic effect of mesenchymal stem cells by regulating the unfolded protein response

    doi: 10.1186/s13036-018-0119-2

    Figure Lengend Snippet: UPR-related gene/protein expression in four groups. a Quantitative RT-PCR for ATF6 , ATF4 , and XBP1 of the four groups in the cultured constructs on 0, 7, 14, and 21 days. These data were normalized to GAPDH . b Western blot results for ATF4, ATF6, XBP1, and COL2A1 at different culture times. COL2A1 was not detectable at 0 day; β-actin was used as a loading control. c Normalized expression of ATF4, ATF6, XBP1, and COL2A1 in response to Western blot analysis. d Immunofluorescence staining of ATF4, ATF6, and XBP1 at 0 days. Scale bar: 50 μm. The values were means ± S.D.; * indicate P

    Article Snippet: The membranes were incubated overnight at 4 °C with appropriate primary antibodies against β-Actin (Boster, Wuhan, China), COL2A1 (Boster), XBP1 (Sigma, USA), ATF6 (Sigma) or ATF4 (Bioss, Beijing, China) after incubation in 1% BSA blocking buffer (Boster) for 1 h at room temperature.

    Techniques: Expressing, Quantitative RT-PCR, Cell Culture, Construct, Western Blot, Immunofluorescence, Staining

    Pioglitazone suppresses RAGE and NF-κB p65 in the brain of STZ-induced diabetic mice. Representative immunoblots of RAGE, NF-κB p65, and β-actin (inner control) were obtained by Western blotting using the appropriate antibodies in the hippocampus and cerebral cortex (A). Quantification of RAGE or NF-κB p65 is expressed as a proportion (in percentage) relative to the control (B and C). Values shown are expressed as the mean±SEM. n =3. b P

    Journal: Acta Pharmacologica Sinica

    Article Title: Pioglitazone ameliorates memory deficits in streptozotocin-induced diabetic mice by reducing brain β-amyloid through PPARγ activation

    doi: 10.1038/aps.2013.11

    Figure Lengend Snippet: Pioglitazone suppresses RAGE and NF-κB p65 in the brain of STZ-induced diabetic mice. Representative immunoblots of RAGE, NF-κB p65, and β-actin (inner control) were obtained by Western blotting using the appropriate antibodies in the hippocampus and cerebral cortex (A). Quantification of RAGE or NF-κB p65 is expressed as a proportion (in percentage) relative to the control (B and C). Values shown are expressed as the mean±SEM. n =3. b P

    Article Snippet: Mouse antibodies were purchased from different companies: anti-Aβ1–42 and anti-receptor for advanced glycation end products (RAGE) were obtained from Abcam Ltd (Hong Kong, China); anti-PPARγ, anti-β-amyloid precursor protein (APP), anti-β-site amyloid precursor protein cleaving enzyme 1 (BACE1), and anti-NF-κB p65 were obtained from Cell Signaling Technology, Inc (Boston, MA, USA); anti-β-actin was obtained from Boster Biotechnology Co, Ltd (Wuhan, China); and secondary antibodies were obtained from Bioworld Technology Co, Ltd (Minneapolis, MN, USA).

    Techniques: Mouse Assay, Western Blot

    Pioglitazone treatment prevents brain Aβ accumulation in STZ-induced diabetic mice. Aβ 1–40 (A) and Aβ 1–42 (B) levels in the hippocampus and cortex were assayed by ELISA. Representative immunoblots of Aβ 1–42 and β-actin (inner control) were obtained by Western blotting using the appropriate antibodies (C), and quantification of Aβ 1–42 is expressed as a proportion (in percentage) relative to the control (D). Values are expressed as the mean±SEM ( n =3–5). b P

    Journal: Acta Pharmacologica Sinica

    Article Title: Pioglitazone ameliorates memory deficits in streptozotocin-induced diabetic mice by reducing brain β-amyloid through PPARγ activation

    doi: 10.1038/aps.2013.11

    Figure Lengend Snippet: Pioglitazone treatment prevents brain Aβ accumulation in STZ-induced diabetic mice. Aβ 1–40 (A) and Aβ 1–42 (B) levels in the hippocampus and cortex were assayed by ELISA. Representative immunoblots of Aβ 1–42 and β-actin (inner control) were obtained by Western blotting using the appropriate antibodies (C), and quantification of Aβ 1–42 is expressed as a proportion (in percentage) relative to the control (D). Values are expressed as the mean±SEM ( n =3–5). b P

    Article Snippet: Mouse antibodies were purchased from different companies: anti-Aβ1–42 and anti-receptor for advanced glycation end products (RAGE) were obtained from Abcam Ltd (Hong Kong, China); anti-PPARγ, anti-β-amyloid precursor protein (APP), anti-β-site amyloid precursor protein cleaving enzyme 1 (BACE1), and anti-NF-κB p65 were obtained from Cell Signaling Technology, Inc (Boston, MA, USA); anti-β-actin was obtained from Boster Biotechnology Co, Ltd (Wuhan, China); and secondary antibodies were obtained from Bioworld Technology Co, Ltd (Minneapolis, MN, USA).

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay, Western Blot

    Pioglitazone treatment reduces brain APP and BACE1 levels in STZ-induced diabetic mice. The expressions of APP and BACE1 in the hippocampus and cortex were detected by Western blotting. Representative immunoblots of APP, BACE1, and β-actin (inner control) are displayed (A), and quantification of APP or BACE1 is expressed as a proportion (in percentage) of the control (B and C). Values shown are expressed as the mean±SEM. n =3. b P

    Journal: Acta Pharmacologica Sinica

    Article Title: Pioglitazone ameliorates memory deficits in streptozotocin-induced diabetic mice by reducing brain β-amyloid through PPARγ activation

    doi: 10.1038/aps.2013.11

    Figure Lengend Snippet: Pioglitazone treatment reduces brain APP and BACE1 levels in STZ-induced diabetic mice. The expressions of APP and BACE1 in the hippocampus and cortex were detected by Western blotting. Representative immunoblots of APP, BACE1, and β-actin (inner control) are displayed (A), and quantification of APP or BACE1 is expressed as a proportion (in percentage) of the control (B and C). Values shown are expressed as the mean±SEM. n =3. b P

    Article Snippet: Mouse antibodies were purchased from different companies: anti-Aβ1–42 and anti-receptor for advanced glycation end products (RAGE) were obtained from Abcam Ltd (Hong Kong, China); anti-PPARγ, anti-β-amyloid precursor protein (APP), anti-β-site amyloid precursor protein cleaving enzyme 1 (BACE1), and anti-NF-κB p65 were obtained from Cell Signaling Technology, Inc (Boston, MA, USA); anti-β-actin was obtained from Boster Biotechnology Co, Ltd (Wuhan, China); and secondary antibodies were obtained from Bioworld Technology Co, Ltd (Minneapolis, MN, USA).

    Techniques: Mouse Assay, Western Blot