apoptosis related human genes  (Qiagen)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    RT² Profiler PCR Array
    Description:
    RT² Profiler PCR Arrays enable quick reliable gene expression analysis of 170 pathways allowing researchers to spend less time at the bench and more time interpreting results The arrays are pathway focused panels of laboratory verified qPCR assays with integrated patented controls to ensure a successful experiment every time PhD trained application specialists are available to provide technical support for the arrays and each array can also be modified to suit unique experimental needs For biomarker discoveries we offer Pathway Plus PCR arrays and all arrays listed below for the Fluidigm BioMark Real Time PCR System
    Catalog Number:
    330231
    Price:
    None
    Category:
    Assay PCR qPCR
    Buy from Supplier


    Structured Review

    Qiagen apoptosis related human genes
    RT² Profiler PCR Array
    RT² Profiler PCR Arrays enable quick reliable gene expression analysis of 170 pathways allowing researchers to spend less time at the bench and more time interpreting results The arrays are pathway focused panels of laboratory verified qPCR assays with integrated patented controls to ensure a successful experiment every time PhD trained application specialists are available to provide technical support for the arrays and each array can also be modified to suit unique experimental needs For biomarker discoveries we offer Pathway Plus PCR arrays and all arrays listed below for the Fluidigm BioMark Real Time PCR System
    https://www.bioz.com/result/apoptosis related human genes/product/Qiagen
    Average 99 stars, based on 5238 article reviews
    Price from $9.99 to $1999.99
    apoptosis related human genes - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Molecular mechanism of action of bisphenol and bisphenol A mediated by oestrogen receptor alpha in growth and apoptosis of breast cancer cells"

    Article Title: Molecular mechanism of action of bisphenol and bisphenol A mediated by oestrogen receptor alpha in growth and apoptosis of breast cancer cells

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.12122

    Differential effect of bisphenol (BP) and bisphenol A (BPA) on growth and apoptosis of ERα positive breast cancer cells. (A) Dose-dependent effects of BP, BPA and (oestradiol) E2 on growth of MCF7 cells treated for 6 days as indicated. The black bar denotes the level of DNA in vehicle treated cells over a 6-day period. The growth is measured as amount of DNA present in each well. (* P
    Figure Legend Snippet: Differential effect of bisphenol (BP) and bisphenol A (BPA) on growth and apoptosis of ERα positive breast cancer cells. (A) Dose-dependent effects of BP, BPA and (oestradiol) E2 on growth of MCF7 cells treated for 6 days as indicated. The black bar denotes the level of DNA in vehicle treated cells over a 6-day period. The growth is measured as amount of DNA present in each well. (* P

    Techniques Used:

    2) Product Images from "CHAC2, downregulated in gastric and colorectal cancers, acted as a tumor suppressor inducing apoptosis and autophagy through unfolded protein response"

    Article Title: CHAC2, downregulated in gastric and colorectal cancers, acted as a tumor suppressor inducing apoptosis and autophagy through unfolded protein response

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2017.405

    Analysis of CHAC2 expression and its degradation pathway in gastric and colorectal cancer cell lines. ( a ) The expression of CHAC2 in gastric and colorectal cancer cell lines was determined by RT-PCR and western bolt, using GAPDH as a control. ( b ) CHAC2 expression in stably transfected cells confirmed by RT-PCR and western blot. ( c ) Evaluation of CHAC2 expression after treatment of 5-FU or BFA. ( d ) Analysis of the potential degradation pathway of CHAC2 using different inhibitors. AGS-CHAC2 and SW620-CHAC2 cells were treated with 20 μ g/ml CHX with or without MG-132 (20 μ M) or Leu (100 μ M) or CQ (50 mM) in a time course. Cell lysates were subjected to western blot with primary antibodies including CHAC2, p-ERK and GAPDH. ( e ) The effect of ubiquitin on CHAC2 expression. AGS-CHAC2 and SW620-CHAC2 cells were incubated with indicated rhHA-ubiquitin for 12 h, and cell lysates were subjected to western blot with primary antibodies including CHAC2, HA tag andGAPDH. ( f ) 12 maximal upregulated genes checked by the Human Ubiquitination Pathway RT 2 Profiler PCR Array were shown in the bar graph. Among these genes, RNF148 expression significantly increased nearly 1863 times in SW620 as compared to SW48. ( g ) The expression of RNF148 in gastric and colorectal cancer cell lines was determined by RT-PCR. ( h ) Evaluation of RNF148 expression after treatment of 5-FU or BFA. 5-FU, 5-fluorouracil; BFA, Brefeldin A; CHX, Cycloheximide; Leu, leupeptin; CQ, Chloroquine; Ubi, rhHA-ubiquitin
    Figure Legend Snippet: Analysis of CHAC2 expression and its degradation pathway in gastric and colorectal cancer cell lines. ( a ) The expression of CHAC2 in gastric and colorectal cancer cell lines was determined by RT-PCR and western bolt, using GAPDH as a control. ( b ) CHAC2 expression in stably transfected cells confirmed by RT-PCR and western blot. ( c ) Evaluation of CHAC2 expression after treatment of 5-FU or BFA. ( d ) Analysis of the potential degradation pathway of CHAC2 using different inhibitors. AGS-CHAC2 and SW620-CHAC2 cells were treated with 20 μ g/ml CHX with or without MG-132 (20 μ M) or Leu (100 μ M) or CQ (50 mM) in a time course. Cell lysates were subjected to western blot with primary antibodies including CHAC2, p-ERK and GAPDH. ( e ) The effect of ubiquitin on CHAC2 expression. AGS-CHAC2 and SW620-CHAC2 cells were incubated with indicated rhHA-ubiquitin for 12 h, and cell lysates were subjected to western blot with primary antibodies including CHAC2, HA tag andGAPDH. ( f ) 12 maximal upregulated genes checked by the Human Ubiquitination Pathway RT 2 Profiler PCR Array were shown in the bar graph. Among these genes, RNF148 expression significantly increased nearly 1863 times in SW620 as compared to SW48. ( g ) The expression of RNF148 in gastric and colorectal cancer cell lines was determined by RT-PCR. ( h ) Evaluation of RNF148 expression after treatment of 5-FU or BFA. 5-FU, 5-fluorouracil; BFA, Brefeldin A; CHX, Cycloheximide; Leu, leupeptin; CQ, Chloroquine; Ubi, rhHA-ubiquitin

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Stable Transfection, Transfection, Incubation, Polymerase Chain Reaction

    3) Product Images from "Effect of ABCG2/BCRP Expression on Efflux and Uptake of Gefitinib in NSCLC Cell Lines"

    Article Title: Effect of ABCG2/BCRP Expression on Efflux and Uptake of Gefitinib in NSCLC Cell Lines

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0141795

    Effect of ABCG2 overexpression on the expression of SLC transporter genes and MPP, glucose and glutamine transport. Down-regulation of SLC transporter expression (SLC22A1, SLC22A3, SLC2A1, SLC38A2, SLC3A2, SLC7A5, and SLC7A6) in HEK293/R2 cells versus HEK293 was evaluated by PCR drug transporter array (A). Fold decrease expression was determined by the ΔΔC T method normalized to five housekeeping genes. Values given are the means (± SD) of three independent experiments (*P
    Figure Legend Snippet: Effect of ABCG2 overexpression on the expression of SLC transporter genes and MPP, glucose and glutamine transport. Down-regulation of SLC transporter expression (SLC22A1, SLC22A3, SLC2A1, SLC38A2, SLC3A2, SLC7A5, and SLC7A6) in HEK293/R2 cells versus HEK293 was evaluated by PCR drug transporter array (A). Fold decrease expression was determined by the ΔΔC T method normalized to five housekeeping genes. Values given are the means (± SD) of three independent experiments (*P

    Techniques Used: Over Expression, Expressing, Polymerase Chain Reaction

    4) Product Images from "Knock-in rats with homozygous PSEN1L435F Alzheimer mutation are viable and show selective γ-secretase activity loss causing low Aβ40/42 and high Aβ43"

    Article Title: Knock-in rats with homozygous PSEN1L435F Alzheimer mutation are viable and show selective γ-secretase activity loss causing low Aβ40/42 and high Aβ43

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.RA120.012542

    RT-PCR analysis of Notch intracellular domain target genes in Psen1 LF rats. A–D , expression levels of several Notch intracellular domain target genes were evaluated in RNA derived from P0 rat brain lysate from Psen1 w/w and Psen1 LF/LF pups. Expression of Cdkn1a ( A ), Cflar ( B ), and Hes1 and Hes5 ( C and D ) showed no significant differences between Psen1 w/w and Psen1 LF/LF rats. Data are represented as mean ± S.D. Data were analyzed by ordinary one-way ANOVA. Samples used: for Cdkn1a , CASP8 , Cfla , and Hes1 , Psen1 w/w n = 3 and Psen1 LF/LF n = 4; for Hes5 , Psen1 w/w n = 6 and Psen1 LF/LF n = 5.
    Figure Legend Snippet: RT-PCR analysis of Notch intracellular domain target genes in Psen1 LF rats. A–D , expression levels of several Notch intracellular domain target genes were evaluated in RNA derived from P0 rat brain lysate from Psen1 w/w and Psen1 LF/LF pups. Expression of Cdkn1a ( A ), Cflar ( B ), and Hes1 and Hes5 ( C and D ) showed no significant differences between Psen1 w/w and Psen1 LF/LF rats. Data are represented as mean ± S.D. Data were analyzed by ordinary one-way ANOVA. Samples used: for Cdkn1a , CASP8 , Cfla , and Hes1 , Psen1 w/w n = 3 and Psen1 LF/LF n = 4; for Hes5 , Psen1 w/w n = 6 and Psen1 LF/LF n = 5.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing, Derivative Assay

    5) Product Images from "Rationalized inhibition of mixed lineage kinase 3 and CD70 enhances life span and antitumor efficacy of CD8+ T cells"

    Article Title: Rationalized inhibition of mixed lineage kinase 3 and CD70 enhances life span and antitumor efficacy of CD8+ T cells

    Journal: Journal for Immunotherapy of Cancer

    doi: 10.1136/jitc-2019-000494

    Loss of MLK3 induces TNFα-TNFRSF1a axis-mediated apoptosis in CD8 + T cells. (A) Experimental approach for RT 2 PCR array for apoptosis-associated genes. (B) RT 2 PCR array for apoptosis-associated genes in purified CD8 + CD38 + T cells derived from WT and MLK3 −/− mice (pooled, n=3 mice/group). (C) Representative histogram plots (upper panel) and quantification (lower panel) of TNFRSF1a surface expression, gated on CD4 + and CD8 + T cells, respectively. Values are the mean±SD, *p
    Figure Legend Snippet: Loss of MLK3 induces TNFα-TNFRSF1a axis-mediated apoptosis in CD8 + T cells. (A) Experimental approach for RT 2 PCR array for apoptosis-associated genes. (B) RT 2 PCR array for apoptosis-associated genes in purified CD8 + CD38 + T cells derived from WT and MLK3 −/− mice (pooled, n=3 mice/group). (C) Representative histogram plots (upper panel) and quantification (lower panel) of TNFRSF1a surface expression, gated on CD4 + and CD8 + T cells, respectively. Values are the mean±SD, *p

    Techniques Used: Polymerase Chain Reaction, Purification, Derivative Assay, Mouse Assay, Expressing

    6) Product Images from "Transient PP2A inhibition alleviates normal tissue stem cell susceptibility to cell death during radiotherapy"

    Article Title: Transient PP2A inhibition alleviates normal tissue stem cell susceptibility to cell death during radiotherapy

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-0559-0

    PP2A inhibition decreases radiosensitivity of stem cells in ex vivo organoid cultures. a Shown are optical microscope images of murine intestinal organoids (upper panel) and murine neurospheres (lower panel) untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Pictures have been taken 48 h after IR treatment. b Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl) or PP2A siRNA (siPP2A). Percentage of cells positive for stem cell marker SSEA1 and apoptotic marker CC-3 was calculated from total SSEA1 positive cells. Scoring was performed 4 h after irradiation. c Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Percentage of cells Lgr5-GFP positive was detected by flow cytometry. Scoring was performed 4 h after irradiation. d Murine hematopoietic stem cells were treated with Cal A or left untreated. Cells were irradiated with 2 Gy and apoptosis analyzed after 16 h by Annexin V labeling. e Human neuroprogenitors were treated with Cal A or left untreated. Cells were irradiated with 6 Gy and apoptosis analyzed after 16 h by Annexin V labeling. Scale bar = 100 μm. Error bars indicate SD; * p
    Figure Legend Snippet: PP2A inhibition decreases radiosensitivity of stem cells in ex vivo organoid cultures. a Shown are optical microscope images of murine intestinal organoids (upper panel) and murine neurospheres (lower panel) untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Pictures have been taken 48 h after IR treatment. b Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl) or PP2A siRNA (siPP2A). Percentage of cells positive for stem cell marker SSEA1 and apoptotic marker CC-3 was calculated from total SSEA1 positive cells. Scoring was performed 4 h after irradiation. c Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Percentage of cells Lgr5-GFP positive was detected by flow cytometry. Scoring was performed 4 h after irradiation. d Murine hematopoietic stem cells were treated with Cal A or left untreated. Cells were irradiated with 2 Gy and apoptosis analyzed after 16 h by Annexin V labeling. e Human neuroprogenitors were treated with Cal A or left untreated. Cells were irradiated with 6 Gy and apoptosis analyzed after 16 h by Annexin V labeling. Scale bar = 100 μm. Error bars indicate SD; * p

    Techniques Used: Inhibition, Ex Vivo, Microscopy, Irradiation, Marker, Flow Cytometry, Cytometry, Labeling

    PP2A suppression influences apoptosis signaling. ES cells were treated with Calyculin A (Cal A) or left untreated. Error bars indicate SD; * p
    Figure Legend Snippet: PP2A suppression influences apoptosis signaling. ES cells were treated with Calyculin A (Cal A) or left untreated. Error bars indicate SD; * p

    Techniques Used:

    7) Product Images from "Thyroid hormone induces progression and invasiveness of squamous cell carcinomas by promoting a ZEB-1/E-cadherin switch"

    Article Title: Thyroid hormone induces progression and invasiveness of squamous cell carcinomas by promoting a ZEB-1/E-cadherin switch

    Journal: Nature Communications

    doi: 10.1038/s41467-019-13140-2

    T3 promotes the EMT of SCC by inducing ZEB-1 transcription. a Vulcano plot of differentially expressed EMT genes measured by real-time PCR in the EMT RT² Profiler™ PCR array (Qiagen). b Heat map of gene expression of EMT markers in normal SCC cells versus D3-depleted SCC cells selected by thresholds of p value
    Figure Legend Snippet: T3 promotes the EMT of SCC by inducing ZEB-1 transcription. a Vulcano plot of differentially expressed EMT genes measured by real-time PCR in the EMT RT² Profiler™ PCR array (Qiagen). b Heat map of gene expression of EMT markers in normal SCC cells versus D3-depleted SCC cells selected by thresholds of p value

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

    Attenuation of TH signaling in D2KO mice enhanced tumor growth and reduced the EMT. a Schematic representation of D2-depletion and the two-step carcinogenesis experiment in 15 CTR and 15 sD2KO samples ( n = 15). b The number of skin lesions counted during DMBA/TPA treatment in sD2KO and CTR mice. c Picture of the dorsal skin from CTR ( n = 15) and sD2KO ( n = 15) mice treated with DMBA/TPA for 20 weeks showing papillomas and advanced SCC (top). H E of the skin lesions from CTR ( n = 8) and sD2KO ( n = 12) mice (bottom). Scale bars represent 200 μm. d The mRNA levels of K6, K8, and E-cadherin/N-cadherin ratio in skin lesions of CTR ( n = 15) and sD2KO ( n = 15) mice measured by real-time PCR analysis. e Immunostaining for K6, K8, vimentin, and E-cadherin was performed on paraffin-embedded sections of dorsal skin lesions ( n = 10 for both groups). Scale bars represent 200 μm. f Schematic representation of D2-depletion from the epidermal compartment at the end of DMBA/TPA treatment (top, n = 15). Representative pictures and H E staining of skin lesions from sD2KO ( n = 12) and CTR ( n = 8) mice treated with DMBA/TPA for 30 weeks (bottom). Scale bars represent 200 μm. g and h The number of tumors (detectable tumors, > 1 mm) and number of large tumors ( > 3 mm) counted during DMBA/TPA treatment in sD2KO and CTR mice. i mRNA levels of K8 in skin lesions of CTR ( n = 15) and sD2KO ( n = 15) mice measured by real-time PCR analysis. * P
    Figure Legend Snippet: Attenuation of TH signaling in D2KO mice enhanced tumor growth and reduced the EMT. a Schematic representation of D2-depletion and the two-step carcinogenesis experiment in 15 CTR and 15 sD2KO samples ( n = 15). b The number of skin lesions counted during DMBA/TPA treatment in sD2KO and CTR mice. c Picture of the dorsal skin from CTR ( n = 15) and sD2KO ( n = 15) mice treated with DMBA/TPA for 20 weeks showing papillomas and advanced SCC (top). H E of the skin lesions from CTR ( n = 8) and sD2KO ( n = 12) mice (bottom). Scale bars represent 200 μm. d The mRNA levels of K6, K8, and E-cadherin/N-cadherin ratio in skin lesions of CTR ( n = 15) and sD2KO ( n = 15) mice measured by real-time PCR analysis. e Immunostaining for K6, K8, vimentin, and E-cadherin was performed on paraffin-embedded sections of dorsal skin lesions ( n = 10 for both groups). Scale bars represent 200 μm. f Schematic representation of D2-depletion from the epidermal compartment at the end of DMBA/TPA treatment (top, n = 15). Representative pictures and H E staining of skin lesions from sD2KO ( n = 12) and CTR ( n = 8) mice treated with DMBA/TPA for 30 weeks (bottom). Scale bars represent 200 μm. g and h The number of tumors (detectable tumors, > 1 mm) and number of large tumors ( > 3 mm) counted during DMBA/TPA treatment in sD2KO and CTR mice. i mRNA levels of K8 in skin lesions of CTR ( n = 15) and sD2KO ( n = 15) mice measured by real-time PCR analysis. * P

    Techniques Used: Mouse Assay, Real-time Polymerase Chain Reaction, Immunostaining, Staining

    8) Product Images from "Expression of p53 Target Genes in the Early Phase of Long-Term Potentiation in the Rat Hippocampal CA1 Area"

    Article Title: Expression of p53 Target Genes in the Early Phase of Long-Term Potentiation in the Rat Hippocampal CA1 Area

    Journal: Neural Plasticity

    doi: 10.1155/2015/242158

    Effect of tetanization on p53, Bax, and Bcl2 in the rat hippocampal CA1 area. (a) Total RNAs were prepared and subjected to real-time PCR for the measurement of mRNAs. The mean of Ct values of five housekeeping genes was used as internal control for normalization as described in Section 2 . (b) Representative Western blots. Whole-cell extracts were prepared from the rat hippocampal CA1 area and subjected to Western blot analysis as described in Section 2 . (c) Relative intensity. The protein bands were analyzed by the computerized densitometric program “Total Lab.” The intensities of the signals were determined from the areas under the curves for each peak and data were graphed. β -actin was used as internal control for normalization. The fold changes were expressed by taking the average value of the group tetanization (−) /nutlin-3 (−) as one. * P
    Figure Legend Snippet: Effect of tetanization on p53, Bax, and Bcl2 in the rat hippocampal CA1 area. (a) Total RNAs were prepared and subjected to real-time PCR for the measurement of mRNAs. The mean of Ct values of five housekeeping genes was used as internal control for normalization as described in Section 2 . (b) Representative Western blots. Whole-cell extracts were prepared from the rat hippocampal CA1 area and subjected to Western blot analysis as described in Section 2 . (c) Relative intensity. The protein bands were analyzed by the computerized densitometric program “Total Lab.” The intensities of the signals were determined from the areas under the curves for each peak and data were graphed. β -actin was used as internal control for normalization. The fold changes were expressed by taking the average value of the group tetanization (−) /nutlin-3 (−) as one. * P

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

    9) Product Images from "GADD45A and CDKN1A are involved in apoptosis and cell cycle modulatory effects of viscumTT with further inactivation of the STAT3 pathway"

    Article Title: GADD45A and CDKN1A are involved in apoptosis and cell cycle modulatory effects of viscumTT with further inactivation of the STAT3 pathway

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-24075-x

    Viscum, TT and viscumTT alter cell cycle-related genes. Expression of cell cycle-related genes of U2OS ( A ), 143B ( B ), Saos-2 ( C ) cells were analyzed by RT² Profiler™ PCR Array after 24 h of viscum, TT and viscumTT treatment. For viscum, mistletoe lectin I (ML) 10 ng/mL, for TT, oleanolic acid (OA) 60 µg/mL and for viscumTT, ML+OA 5 ng/mL + 50 µg/mL were used. Array was performed once for each cell line and the fold-regulation cut-off was set to 2 and the p-value cut off was set to p ≤ 0.05 by the software.
    Figure Legend Snippet: Viscum, TT and viscumTT alter cell cycle-related genes. Expression of cell cycle-related genes of U2OS ( A ), 143B ( B ), Saos-2 ( C ) cells were analyzed by RT² Profiler™ PCR Array after 24 h of viscum, TT and viscumTT treatment. For viscum, mistletoe lectin I (ML) 10 ng/mL, for TT, oleanolic acid (OA) 60 µg/mL and for viscumTT, ML+OA 5 ng/mL + 50 µg/mL were used. Array was performed once for each cell line and the fold-regulation cut-off was set to 2 and the p-value cut off was set to p ≤ 0.05 by the software.

    Techniques Used: Expressing, Polymerase Chain Reaction, Software

    10) Product Images from "Adipocyte arrestin domain-containing 3 protein (Arrdc3) regulates uncoupling protein 1 (Ucp1) expression in white adipose independently of canonical changes in β-adrenergic receptor signaling"

    Article Title: Adipocyte arrestin domain-containing 3 protein (Arrdc3) regulates uncoupling protein 1 (Ucp1) expression in white adipose independently of canonical changes in β-adrenergic receptor signaling

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0173823

    Arrdc3 deletion increases PPAR target gene expression in adipocytes. Quantitative PCR array analysis of gene expression of PPAR target genes, PPAR cofactors and PPARs and associated transcription factors in control versus Arrdc3 SVF-derived adipocytes in vitro (n = 3 mice per group). Only significantly different genes are displayed. *p≤ 0.05.
    Figure Legend Snippet: Arrdc3 deletion increases PPAR target gene expression in adipocytes. Quantitative PCR array analysis of gene expression of PPAR target genes, PPAR cofactors and PPARs and associated transcription factors in control versus Arrdc3 SVF-derived adipocytes in vitro (n = 3 mice per group). Only significantly different genes are displayed. *p≤ 0.05.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Derivative Assay, In Vitro, Mouse Assay

    11) Product Images from "Transient PP2A inhibition alleviates normal tissue stem cell susceptibility to cell death during radiotherapy"

    Article Title: Transient PP2A inhibition alleviates normal tissue stem cell susceptibility to cell death during radiotherapy

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-0559-0

    PP2A inhibition decreases radiosensitivity of stem cells in ex vivo organoid cultures. a Shown are optical microscope images of murine intestinal organoids (upper panel) and murine neurospheres (lower panel) untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Pictures have been taken 48 h after IR treatment. b Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl) or PP2A siRNA (siPP2A). Percentage of cells positive for stem cell marker SSEA1 and apoptotic marker CC-3 was calculated from total SSEA1 positive cells. Scoring was performed 4 h after irradiation. c Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Percentage of cells Lgr5-GFP positive was detected by flow cytometry. Scoring was performed 4 h after irradiation. d Murine hematopoietic stem cells were treated with Cal A or left untreated. Cells were irradiated with 2 Gy and apoptosis analyzed after 16 h by Annexin V labeling. e Human neuroprogenitors were treated with Cal A or left untreated. Cells were irradiated with 6 Gy and apoptosis analyzed after 16 h by Annexin V labeling. Scale bar = 100 μm. Error bars indicate SD; * p
    Figure Legend Snippet: PP2A inhibition decreases radiosensitivity of stem cells in ex vivo organoid cultures. a Shown are optical microscope images of murine intestinal organoids (upper panel) and murine neurospheres (lower panel) untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Pictures have been taken 48 h after IR treatment. b Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl) or PP2A siRNA (siPP2A). Percentage of cells positive for stem cell marker SSEA1 and apoptotic marker CC-3 was calculated from total SSEA1 positive cells. Scoring was performed 4 h after irradiation. c Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Percentage of cells Lgr5-GFP positive was detected by flow cytometry. Scoring was performed 4 h after irradiation. d Murine hematopoietic stem cells were treated with Cal A or left untreated. Cells were irradiated with 2 Gy and apoptosis analyzed after 16 h by Annexin V labeling. e Human neuroprogenitors were treated with Cal A or left untreated. Cells were irradiated with 6 Gy and apoptosis analyzed after 16 h by Annexin V labeling. Scale bar = 100 μm. Error bars indicate SD; * p

    Techniques Used: Inhibition, Ex Vivo, Microscopy, Irradiation, Marker, Flow Cytometry, Cytometry, Labeling

    12) Product Images from "Extracellular Vesicles–Encapsulated MicroRNA-125b Produced in Genetically Modified Mesenchymal Stromal Cells Inhibits Hepatocellular Carcinoma Cell Proliferation"

    Article Title: Extracellular Vesicles–Encapsulated MicroRNA-125b Produced in Genetically Modified Mesenchymal Stromal Cells Inhibits Hepatocellular Carcinoma Cell Proliferation

    Journal: Cells

    doi: 10.3390/cells8121560

    Delivery of EV isolated from adipose tissue derived stromal cells genetically modified with ExoMotif-tagged microRNA-125b affects p53 expression. ( a ) Immunoblot analysis for p53 performed on Hep G2 and HCT 116 cells cultured in presence (+) of miRNA-125b containing or control (−) EVs for 144 h; GAPDH protein level was used as loading control. Densitometry analysis performed using ImageJ software is indicated. ( b ) Analysis of the p53 signaling pathway using a specific RT2 Profiler PCR Array. The expression analysis of 84 genes in EV miR-125b group and Ctrl group was performed. Scatter plot comparison shows gene expression changes in the two groups. The red and green dots represent up-regulated and down-regulated genes, respectively. ( c ) Table reporting the genes changed at least a two-fold differential expression in EV miR-125b group against the control group.
    Figure Legend Snippet: Delivery of EV isolated from adipose tissue derived stromal cells genetically modified with ExoMotif-tagged microRNA-125b affects p53 expression. ( a ) Immunoblot analysis for p53 performed on Hep G2 and HCT 116 cells cultured in presence (+) of miRNA-125b containing or control (−) EVs for 144 h; GAPDH protein level was used as loading control. Densitometry analysis performed using ImageJ software is indicated. ( b ) Analysis of the p53 signaling pathway using a specific RT2 Profiler PCR Array. The expression analysis of 84 genes in EV miR-125b group and Ctrl group was performed. Scatter plot comparison shows gene expression changes in the two groups. The red and green dots represent up-regulated and down-regulated genes, respectively. ( c ) Table reporting the genes changed at least a two-fold differential expression in EV miR-125b group against the control group.

    Techniques Used: Isolation, Derivative Assay, Genetically Modified, Expressing, Cell Culture, Software, Polymerase Chain Reaction

    13) Product Images from "Doxycycline Enhances Survival and Self-Renewal of Human Pluripotent Stem Cells"

    Article Title: Doxycycline Enhances Survival and Self-Renewal of Human Pluripotent Stem Cells

    Journal: Stem Cell Reports

    doi: 10.1016/j.stemcr.2014.06.013

    Practical Benefits of Doxycycline Supplementation in hESC/iPSC Cultures The effects of doxycycline supplementation were examined in undifferentiated hESC/iPSC cultures maintained in clusters without dissociation on MEF feeder cells. (A and B) FACS analyses for determining the fraction of cells undergoing apoptosis (A) and the fraction that had accumulated in each cell-cycle phase (B). Small clusters of H9 hESCs or Lenti-1 hiPSCs (10-50 cells) were plated and cultured in the presence or absence of doxycycline (1 μg/ml) for 6 days. The hESC/iPSC clusters were mechanically harvested, dissociated using TrypLE (A) or Accutase (B), 2 hr prior to Annexin V/PI (A) or PI (B) staining. The stained populations were analyzed using FACS. n = 2 or 3 technical replicates/experiment, n = 2 experiments (A). n = 3 experiments (B) ∗ p
    Figure Legend Snippet: Practical Benefits of Doxycycline Supplementation in hESC/iPSC Cultures The effects of doxycycline supplementation were examined in undifferentiated hESC/iPSC cultures maintained in clusters without dissociation on MEF feeder cells. (A and B) FACS analyses for determining the fraction of cells undergoing apoptosis (A) and the fraction that had accumulated in each cell-cycle phase (B). Small clusters of H9 hESCs or Lenti-1 hiPSCs (10-50 cells) were plated and cultured in the presence or absence of doxycycline (1 μg/ml) for 6 days. The hESC/iPSC clusters were mechanically harvested, dissociated using TrypLE (A) or Accutase (B), 2 hr prior to Annexin V/PI (A) or PI (B) staining. The stained populations were analyzed using FACS. n = 2 or 3 technical replicates/experiment, n = 2 experiments (A). n = 3 experiments (B) ∗ p

    Techniques Used: FACS, Cell Culture, Staining

    Doxycycline Permits Cell Survival and Maintenance of Undifferentiated hESCs through PI3K-AKT Signal Activation. (A–C) Cell dissociation-induced actomyosin hyperactivation is prevented by Y-27632, but not by doxycycline treatment. H9 hESCs were incubated with doxycycline or Y-27632 or without these chemicals (control) for 1 hr prior to cell dissociation. Cells were then harvested 0, 10, and 30 min after cell dissociation and subjected to western blot analyses to detect proteins involved in cell-dissociation-induced apoptosis (A). Shown in (B) and (C) are representative pMLC-immunostained and phase-contrast images of dissociated hESCs in the cultures treated with Y-27632 or doxycycline. Cells pretreated with Y-27632 or doxycycline were dissociated, directly plated onto Matrigel with mTeSR1 and cultured in the continuous presence of the chemicals. The images were captured at the indicated time after plating (dissociation). Scale bars, 30 μm. (D) Estimation of intracellular signals activated by doxycycline. H9 hESCs were incubated without (control) or with doxycycline (1 μg/ml) for 30 min before harvesting and then subjected to immunoblot analyses using the human phosphokinase blot array, which is designed to detect 46 phosphorylated intracellular proteins. The array analyses were performed in technical triplicate. Shown is a representative pair of blots of the untreated control and doxycycline-treated cells. In the blots, spots whose intensities are greater ( > 1.5-fold) in the doxycycline-treated group, compared to the control, are marked with dotted circles and listed with the fold increase (parentheses) in the inset. ∗∗ p
    Figure Legend Snippet: Doxycycline Permits Cell Survival and Maintenance of Undifferentiated hESCs through PI3K-AKT Signal Activation. (A–C) Cell dissociation-induced actomyosin hyperactivation is prevented by Y-27632, but not by doxycycline treatment. H9 hESCs were incubated with doxycycline or Y-27632 or without these chemicals (control) for 1 hr prior to cell dissociation. Cells were then harvested 0, 10, and 30 min after cell dissociation and subjected to western blot analyses to detect proteins involved in cell-dissociation-induced apoptosis (A). Shown in (B) and (C) are representative pMLC-immunostained and phase-contrast images of dissociated hESCs in the cultures treated with Y-27632 or doxycycline. Cells pretreated with Y-27632 or doxycycline were dissociated, directly plated onto Matrigel with mTeSR1 and cultured in the continuous presence of the chemicals. The images were captured at the indicated time after plating (dissociation). Scale bars, 30 μm. (D) Estimation of intracellular signals activated by doxycycline. H9 hESCs were incubated without (control) or with doxycycline (1 μg/ml) for 30 min before harvesting and then subjected to immunoblot analyses using the human phosphokinase blot array, which is designed to detect 46 phosphorylated intracellular proteins. The array analyses were performed in technical triplicate. Shown is a representative pair of blots of the untreated control and doxycycline-treated cells. In the blots, spots whose intensities are greater ( > 1.5-fold) in the doxycycline-treated group, compared to the control, are marked with dotted circles and listed with the fold increase (parentheses) in the inset. ∗∗ p

    Techniques Used: Activation Assay, Incubation, Western Blot, Cell Culture

    14) Product Images from "Kallikrein-related peptidases 4, 5, 6 and 7 regulate tumour-associated factors in serous ovarian cancer"

    Article Title: Kallikrein-related peptidases 4, 5, 6 and 7 regulate tumour-associated factors in serous ovarian cancer

    Journal: British Journal of Cancer

    doi: 10.1038/s41416-018-0260-1

    Analysis of mRNA expression in OV-KLK4–7 versus control OV-VC cells via PCR array and qPCR. Selected genes identified as being deregulated in OV-KLK4–7 (red columns) versus OV-VC (blue columns) cells in the PCR array were validated by qPCR using independently isolated RNA. OV-KLK4–7 cells displayed significant upregulation of MSN ( a ), KRT19 ( b ), COL5A2 ( c ), COL1A2 ( d ), BMP5 ( e ) and F10 ( f ) as well as downregulation of KRT7 ( g ), JUNB ( h ), BMP4 ( j ) and MMP1 ( k ) compared to OV-VC cells (* P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001)
    Figure Legend Snippet: Analysis of mRNA expression in OV-KLK4–7 versus control OV-VC cells via PCR array and qPCR. Selected genes identified as being deregulated in OV-KLK4–7 (red columns) versus OV-VC (blue columns) cells in the PCR array were validated by qPCR using independently isolated RNA. OV-KLK4–7 cells displayed significant upregulation of MSN ( a ), KRT19 ( b ), COL5A2 ( c ), COL1A2 ( d ), BMP5 ( e ) and F10 ( f ) as well as downregulation of KRT7 ( g ), JUNB ( h ), BMP4 ( j ) and MMP1 ( k ) compared to OV-VC cells (* P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001)

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

    15) Product Images from "Interplay between MycN and c-Myc regulates radioresistance and cancer stem cell phenotype in neuroblastoma upon glutamine deprivation"

    Article Title: Interplay between MycN and c-Myc regulates radioresistance and cancer stem cell phenotype in neuroblastoma upon glutamine deprivation

    Journal: Theranostics

    doi: 10.7150/thno.42602

    Glutamine deprivation modifies DNA repair gene pathway and redox balance in neuroblastoma cells. (A) Expression levels of 84 DNA repair genes analyzed using DNA RT² Profiler PCR Array Human DNA Repair in BE(2)-C cells upon 24 h of glutamine deprivation. Data are from two pooled experiments. (B, C) Correlation of mRNA expression for (B) 17 genes downregulated upon glutamine deprivation and (C) 16 genes upregulated upon glutamine starvation with c-MYC and MYCN genes in the neuroblastoma TARGET patient cohort. (D, E) Correlation map for (D) 6 clinically relevant genes, which were downregulated upon glutamine deprivation and (E) 4 clinically relevant genes, which were upregulated upon glutamine deprivation, with c-MYC and MYCN . (F) Representative western blot of total Chk1 and p-Chk1 levels in BE(2)-C cells upon glutamine deprivation and irradiation (4 Gy). GAPDH was used as a loading control. (G) Relative mRNA expression of CHK1 , MYCN , c-MYC and 6 DNA repair genes upon glutamine deprivation in BE(2)-C cells. (H) Analysis of ROS levels in SH-SY5Y (red) and BE(2)-C (blue) cells upon 48 h glutamine deprivation and 4 Gy X-rays. NAC and antimycin A used as negative and positive controls. Data are reported as averages relative to control (n = 3; ± S.E.M; *p
    Figure Legend Snippet: Glutamine deprivation modifies DNA repair gene pathway and redox balance in neuroblastoma cells. (A) Expression levels of 84 DNA repair genes analyzed using DNA RT² Profiler PCR Array Human DNA Repair in BE(2)-C cells upon 24 h of glutamine deprivation. Data are from two pooled experiments. (B, C) Correlation of mRNA expression for (B) 17 genes downregulated upon glutamine deprivation and (C) 16 genes upregulated upon glutamine starvation with c-MYC and MYCN genes in the neuroblastoma TARGET patient cohort. (D, E) Correlation map for (D) 6 clinically relevant genes, which were downregulated upon glutamine deprivation and (E) 4 clinically relevant genes, which were upregulated upon glutamine deprivation, with c-MYC and MYCN . (F) Representative western blot of total Chk1 and p-Chk1 levels in BE(2)-C cells upon glutamine deprivation and irradiation (4 Gy). GAPDH was used as a loading control. (G) Relative mRNA expression of CHK1 , MYCN , c-MYC and 6 DNA repair genes upon glutamine deprivation in BE(2)-C cells. (H) Analysis of ROS levels in SH-SY5Y (red) and BE(2)-C (blue) cells upon 48 h glutamine deprivation and 4 Gy X-rays. NAC and antimycin A used as negative and positive controls. Data are reported as averages relative to control (n = 3; ± S.E.M; *p

    Techniques Used: Expressing, Polymerase Chain Reaction, Western Blot, Irradiation

    Glutamine deprivation modulates CSC properties in neuroblastoma cells. (A) Expression of 84 CSC-related genes analyzed by RT² Profiler PCR Array Human Cancer Stem Cells in BE(2)-C and SH-SY5Y cells upon 24 h of glutamine deprivation. Data are from two pooled experiments. (B) Relative expression level for PROM1 following c-MYC or MYCN knockdown in BE(2)-C and SH-SY5Y cells. Data are from three pooled experiments. (C) Flow cytometry analysis of CD133-positive populations in BE(2)-C and SH-SY5Y cells upon glutamine deprivation (n ≥ 3; ± S.E.M; *p
    Figure Legend Snippet: Glutamine deprivation modulates CSC properties in neuroblastoma cells. (A) Expression of 84 CSC-related genes analyzed by RT² Profiler PCR Array Human Cancer Stem Cells in BE(2)-C and SH-SY5Y cells upon 24 h of glutamine deprivation. Data are from two pooled experiments. (B) Relative expression level for PROM1 following c-MYC or MYCN knockdown in BE(2)-C and SH-SY5Y cells. Data are from three pooled experiments. (C) Flow cytometry analysis of CD133-positive populations in BE(2)-C and SH-SY5Y cells upon glutamine deprivation (n ≥ 3; ± S.E.M; *p

    Techniques Used: Expressing, Polymerase Chain Reaction, Flow Cytometry

    16) Product Images from "Gold Nanocomplex Strongly Modulates the PI3K/Akt Pathway and Other Pathways in MCF-7 Breast Cancer Cell Line"

    Article Title: Gold Nanocomplex Strongly Modulates the PI3K/Akt Pathway and Other Pathways in MCF-7 Breast Cancer Cell Line

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21093320

    PCR array analysis of PI3K/Akt signaling pathway of MCF-7 cells. A representative heat map of expression values of genes involved in PI3K pathways among different treated groups; the nanocomplex, the free drug and gold nanorods (GNR) only (presented in the map as A: nanocomplex; D: the free drug and G: GNR) compared to their control untreated cells.
    Figure Legend Snippet: PCR array analysis of PI3K/Akt signaling pathway of MCF-7 cells. A representative heat map of expression values of genes involved in PI3K pathways among different treated groups; the nanocomplex, the free drug and gold nanorods (GNR) only (presented in the map as A: nanocomplex; D: the free drug and G: GNR) compared to their control untreated cells.

    Techniques Used: Polymerase Chain Reaction, Expressing

    17) Product Images from "Mouse homolog of the human TP53 R337H mutation reveals its role in tumorigenesis"

    Article Title: Mouse homolog of the human TP53 R337H mutation reveals its role in tumorigenesis

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-18-0016

    Liver tissues of p53 334H/H mice exposed to DEN show increased DNA damage response markers but diminished p53 activity. A , liver tissue samples from untreated (control) and DEN-treated (24 h) mice were immunoblotted with the indicated antibodies. Phospho-p53 (S15) and histone γ-H2AX (S139) serve as DNA damage response markers. Each lane represents an independent replicate mouse of untreated control and DEN-treated conditions. p53 −/− mouse liver serves as negative control for p53 protein and activity. Arrowheads indicate p53-dependent p21 induced by DEN; note absence of this band in the p53 −/− lane. B , apoptosis imaging and quantification in liver sections before or after DEN treatment. Fluorescent images of TUNEL stained sections of the indicated p53 genotype are shown. The fractions of apoptotic TUNEL-positive nuclei (green) versus total nuclei stained with Hoechst 33342 (blue) were determined ( n = 3). Scale bar = 50 µm. C , FACS cell cycle analysis of untreated control and DEN-treated liver cells stained with propidium iodide. The relative fraction of cells in the different cell cycle stages (G1, S and G2/M) were quantified ( n = 5). p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); and homozygous mutant ( H/H ). Values are mean ± SD. * P
    Figure Legend Snippet: Liver tissues of p53 334H/H mice exposed to DEN show increased DNA damage response markers but diminished p53 activity. A , liver tissue samples from untreated (control) and DEN-treated (24 h) mice were immunoblotted with the indicated antibodies. Phospho-p53 (S15) and histone γ-H2AX (S139) serve as DNA damage response markers. Each lane represents an independent replicate mouse of untreated control and DEN-treated conditions. p53 −/− mouse liver serves as negative control for p53 protein and activity. Arrowheads indicate p53-dependent p21 induced by DEN; note absence of this band in the p53 −/− lane. B , apoptosis imaging and quantification in liver sections before or after DEN treatment. Fluorescent images of TUNEL stained sections of the indicated p53 genotype are shown. The fractions of apoptotic TUNEL-positive nuclei (green) versus total nuclei stained with Hoechst 33342 (blue) were determined ( n = 3). Scale bar = 50 µm. C , FACS cell cycle analysis of untreated control and DEN-treated liver cells stained with propidium iodide. The relative fraction of cells in the different cell cycle stages (G1, S and G2/M) were quantified ( n = 5). p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); and homozygous mutant ( H/H ). Values are mean ± SD. * P

    Techniques Used: Mouse Assay, Activity Assay, Negative Control, Imaging, TUNEL Assay, Staining, FACS, Cell Cycle Assay, Mutagenesis

    p53 R334H does not significantly affect lifespan or cancer incidence in mice. A , Kaplan-Meier survival plot of male and female p53 334R/R , p53 334R/H , and p53 334H/H mice; the respective median survival times (male/female) for these 3 genotypes were 114/99, 103/103, and 103/101 wk. No significant difference in survival time was observed by log-rank test ( n = 9–29). B , incidence of cancer by p53 R334 genotype at necropsy after reaching survival endpoint ( n = 24–39). C , spectrum of cancer diagnosis by p53 R334 genotype ( n = 24–39). p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); and homozygous mutant ( H/H ). Both incidence and spectrum of cancer were not significantly different compared with wild-type mice by Fisher’s exact test.
    Figure Legend Snippet: p53 R334H does not significantly affect lifespan or cancer incidence in mice. A , Kaplan-Meier survival plot of male and female p53 334R/R , p53 334R/H , and p53 334H/H mice; the respective median survival times (male/female) for these 3 genotypes were 114/99, 103/103, and 103/101 wk. No significant difference in survival time was observed by log-rank test ( n = 9–29). B , incidence of cancer by p53 R334 genotype at necropsy after reaching survival endpoint ( n = 24–39). C , spectrum of cancer diagnosis by p53 R334 genotype ( n = 24–39). p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); and homozygous mutant ( H/H ). Both incidence and spectrum of cancer were not significantly different compared with wild-type mice by Fisher’s exact test.

    Techniques Used: Mouse Assay, Mutagenesis

    p53 R334H confers increased susceptibility to DEN-induced liver tumorigenesis in mice. A , representative images showing liver tumor formation 42 wk after DEN treatment (top panel). Small tumor indicated by arrowhead. Lower panels show representative H E stained photomicrographs of liver tumor sections under low (5×) and high (40×, boxed areas in 5×) objective magnification. Broken lines delineate the following regions of histopathology: normal (NL), hyperplasia (HP), adenoma (AD), and carcinoma (CA). Scale bar = 500 µm and 50 µm. B , quantification of tumors visible on the surface of the liver by size (≥ 1 mm diameter) ( n = 14–20). C , liver weight was normalized by body weight to provide an index of tumor burden ( n = 14–20). D , quantification of liver tumor histopathologic diagnosis as abbreviated in (A). Counts indicate the number of specific diagnosis made in each liver sample ( n = 5). p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); and homozygous mutant ( H/H ). Values are mean ± SD. Statistical differences were tested by 1-way ANOVA. * P
    Figure Legend Snippet: p53 R334H confers increased susceptibility to DEN-induced liver tumorigenesis in mice. A , representative images showing liver tumor formation 42 wk after DEN treatment (top panel). Small tumor indicated by arrowhead. Lower panels show representative H E stained photomicrographs of liver tumor sections under low (5×) and high (40×, boxed areas in 5×) objective magnification. Broken lines delineate the following regions of histopathology: normal (NL), hyperplasia (HP), adenoma (AD), and carcinoma (CA). Scale bar = 500 µm and 50 µm. B , quantification of tumors visible on the surface of the liver by size (≥ 1 mm diameter) ( n = 14–20). C , liver weight was normalized by body weight to provide an index of tumor burden ( n = 14–20). D , quantification of liver tumor histopathologic diagnosis as abbreviated in (A). Counts indicate the number of specific diagnosis made in each liver sample ( n = 5). p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); and homozygous mutant ( H/H ). Values are mean ± SD. Statistical differences were tested by 1-way ANOVA. * P

    Techniques Used: Mouse Assay, Staining, Histopathology, Mutagenesis

    Partial loss of p53 transcriptional activity in liver tissue of homozygous mutant p53 334H/H mice treated with DEN. A , color-coded relative expression of 84 p53-regulated genes in untreated (control) and DEN-treated (24 h) mouse livers of the indicated p53 genotype analyzed by RT-PCR screening array ( n = 3). Red and green indicate induced and repressed mRNA levels, respectively. Expression levels were normalized to control R/R samples. B , the screening array was filtered for genes significantly induced (≥ 2-fold) in wild-type mice by DEN treatment. Of the 11 genes that were identified, 10 were significantly lower in the homozygous mutant compared with wild-type livers ( n = 3). p21, DR5, WIG, NOXA, and PUMA are referred to as CDKN1A, TNFRSF10B, ZMAT3, PMAIP1, and BBC3, respectively, in (A). C , the mRNA expression levels of 4 prototypical p53-regulated genes were confirmed by independent real-time RT-PCR ( n = 4). Expression levels were normalized relative to wild-type samples. p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); homozygous mutant ( H/H ); and p53 null ( −/− ). Values are mean ± SD. Statistical differences were tested by 1-way ANOVA. * P
    Figure Legend Snippet: Partial loss of p53 transcriptional activity in liver tissue of homozygous mutant p53 334H/H mice treated with DEN. A , color-coded relative expression of 84 p53-regulated genes in untreated (control) and DEN-treated (24 h) mouse livers of the indicated p53 genotype analyzed by RT-PCR screening array ( n = 3). Red and green indicate induced and repressed mRNA levels, respectively. Expression levels were normalized to control R/R samples. B , the screening array was filtered for genes significantly induced (≥ 2-fold) in wild-type mice by DEN treatment. Of the 11 genes that were identified, 10 were significantly lower in the homozygous mutant compared with wild-type livers ( n = 3). p21, DR5, WIG, NOXA, and PUMA are referred to as CDKN1A, TNFRSF10B, ZMAT3, PMAIP1, and BBC3, respectively, in (A). C , the mRNA expression levels of 4 prototypical p53-regulated genes were confirmed by independent real-time RT-PCR ( n = 4). Expression levels were normalized relative to wild-type samples. p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); homozygous mutant ( H/H ); and p53 null ( −/− ). Values are mean ± SD. Statistical differences were tested by 1-way ANOVA. * P

    Techniques Used: Activity Assay, Mutagenesis, Mouse Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

    p53 oligomerization is decreased in p53 334H/H mouse tissues. A , liver tissue lysates were cross-linked with glutaraldehyde, resolved in SDS-PAGE gel, and immunoblotted. Note p53 antibody specificity demonstrated by lack of immunoreactivity in the p53 −/− sample. Protein standards are in kD. Triplicate lanes of each genotype represent liver samples from 3 separate mice. B, fraction of p53 oligomers (T, tetramer; D, dimer; M, monomer) within each lane of immunoblot (A) quantified by densitometry and compared with the respective wild-type oligomer ( n = 3). C , p53 immunoblot of cross-linked liver lysates obtained from mice 6 h after p53 induction by doxorubicin treatment. Duplicate lanes of each genotype represent liver samples from 2 separate mice. D , p53 binding to the p53 response element (p53RE) of p21 in γ-irradiated (γ-IR) mouse liver. ChIP was performed using nonspecific IgG or anti-p53 antibody. p53RE binding is shown relative to wild-type non-specific IgG samples ( n = 3). E, induction of p21 mRNA in the indicated tissues by γ-IR quantified by RT-PCR ( n = 3). Levels were normalized relative to a housekeeping gene TIF . Bone marrow (BM); small intestine. p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); homozygous mutant ( H/H ); and p53 null ( −/− ). Values are mean ± SD. Compared to wild-type samples within each group, statistical differences were tested by 1-way ANOVA. * P
    Figure Legend Snippet: p53 oligomerization is decreased in p53 334H/H mouse tissues. A , liver tissue lysates were cross-linked with glutaraldehyde, resolved in SDS-PAGE gel, and immunoblotted. Note p53 antibody specificity demonstrated by lack of immunoreactivity in the p53 −/− sample. Protein standards are in kD. Triplicate lanes of each genotype represent liver samples from 3 separate mice. B, fraction of p53 oligomers (T, tetramer; D, dimer; M, monomer) within each lane of immunoblot (A) quantified by densitometry and compared with the respective wild-type oligomer ( n = 3). C , p53 immunoblot of cross-linked liver lysates obtained from mice 6 h after p53 induction by doxorubicin treatment. Duplicate lanes of each genotype represent liver samples from 2 separate mice. D , p53 binding to the p53 response element (p53RE) of p21 in γ-irradiated (γ-IR) mouse liver. ChIP was performed using nonspecific IgG or anti-p53 antibody. p53RE binding is shown relative to wild-type non-specific IgG samples ( n = 3). E, induction of p21 mRNA in the indicated tissues by γ-IR quantified by RT-PCR ( n = 3). Levels were normalized relative to a housekeeping gene TIF . Bone marrow (BM); small intestine. p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); homozygous mutant ( H/H ); and p53 null ( −/− ). Values are mean ± SD. Compared to wild-type samples within each group, statistical differences were tested by 1-way ANOVA. * P

    Techniques Used: SDS Page, Mouse Assay, Binding Assay, Irradiation, Chromatin Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction, Mutagenesis

    p53 R334H mutation knockin construct and screening strategy. A , conventional ES cell-mediated strategy was used to knockin the c.1001G > A missense mutation into the p53 gene ( p53 334H ; mouse R334H corresponding to human R337H). Mice with germline transmission of the mutation were crossed with cre-expressing mice to remove the neomycin (Neo) resistance gene flanked by lox P sites (triangles). Asterisk indicates the mutated nucleotide position in exon 10 and horizontal arrowheads indicate forward and reverse primers for genotype screening. B , sequencing of the complementary strand confirmed correct integration of the mutation. The wild-type (WT) and mutant (Mut) codon positions are underlined. C , genomic DNA PCR was also performed to confirm the genotypes of mice. The patterns of WT (305 bp) and Mut (611 bp) PCR products indicate wild-type ( R/R ), heterozygous mutant ( R/H ), and homozygous mutant ( H/H ) genotypes.
    Figure Legend Snippet: p53 R334H mutation knockin construct and screening strategy. A , conventional ES cell-mediated strategy was used to knockin the c.1001G > A missense mutation into the p53 gene ( p53 334H ; mouse R334H corresponding to human R337H). Mice with germline transmission of the mutation were crossed with cre-expressing mice to remove the neomycin (Neo) resistance gene flanked by lox P sites (triangles). Asterisk indicates the mutated nucleotide position in exon 10 and horizontal arrowheads indicate forward and reverse primers for genotype screening. B , sequencing of the complementary strand confirmed correct integration of the mutation. The wild-type (WT) and mutant (Mut) codon positions are underlined. C , genomic DNA PCR was also performed to confirm the genotypes of mice. The patterns of WT (305 bp) and Mut (611 bp) PCR products indicate wild-type ( R/R ), heterozygous mutant ( R/H ), and homozygous mutant ( H/H ) genotypes.

    Techniques Used: Mutagenesis, Knock-In, Construct, Mouse Assay, Transmission Assay, Expressing, Sequencing, Polymerase Chain Reaction

    18) Product Images from "Preconditioning by Hydrogen Peroxide Enhances Multiple Properties of Human Decidua Basalis Mesenchymal Stem/Multipotent Stromal Cells"

    Article Title: Preconditioning by Hydrogen Peroxide Enhances Multiple Properties of Human Decidua Basalis Mesenchymal Stem/Multipotent Stromal Cells

    Journal: Stem Cells International

    doi: 10.1155/2018/6480793

    List of stress-related genes modulated in DBMSCs after exposure to H2 O2 .
    Figure Legend Snippet: List of stress-related genes modulated in DBMSCs after exposure to H2 O2 .

    Techniques Used:

    19) Product Images from "GADD45A and CDKN1A are involved in apoptosis and cell cycle modulatory effects of viscumTT with further inactivation of the STAT3 pathway"

    Article Title: GADD45A and CDKN1A are involved in apoptosis and cell cycle modulatory effects of viscumTT with further inactivation of the STAT3 pathway

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-24075-x

    Viscum, TT and viscumTT alter cell cycle-related genes. Expression of cell cycle-related genes of U2OS ( A ), 143B ( B ), Saos-2 ( C ) cells were analyzed by RT² Profiler™ PCR Array after 24 h of viscum, TT and viscumTT treatment. For viscum, mistletoe lectin I (ML) 10 ng/mL, for TT, oleanolic acid (OA) 60 µg/mL and for viscumTT, ML+OA 5 ng/mL + 50 µg/mL were used. Array was performed once for each cell line and the fold-regulation cut-off was set to 2 and the p-value cut off was set to p ≤ 0.05 by the software.
    Figure Legend Snippet: Viscum, TT and viscumTT alter cell cycle-related genes. Expression of cell cycle-related genes of U2OS ( A ), 143B ( B ), Saos-2 ( C ) cells were analyzed by RT² Profiler™ PCR Array after 24 h of viscum, TT and viscumTT treatment. For viscum, mistletoe lectin I (ML) 10 ng/mL, for TT, oleanolic acid (OA) 60 µg/mL and for viscumTT, ML+OA 5 ng/mL + 50 µg/mL were used. Array was performed once for each cell line and the fold-regulation cut-off was set to 2 and the p-value cut off was set to p ≤ 0.05 by the software.

    Techniques Used: Expressing, Polymerase Chain Reaction, Software

    20) Product Images from "Mouse homolog of the human TP53 R337H mutation reveals its role in tumorigenesis"

    Article Title: Mouse homolog of the human TP53 R337H mutation reveals its role in tumorigenesis

    Journal: Cancer research

    doi: 10.1158/0008-5472.CAN-18-0016

    Partial loss of p53 transcriptional activity in liver tissue of homozygous mutant p53 334H/H mice treated with DEN. A , color-coded relative expression of 84 p53-regulated genes in untreated (control) and DEN-treated (24 h) mouse livers of the indicated p53 genotype analyzed by RT-PCR screening array ( n = 3). Red and green indicate induced and repressed mRNA levels, respectively. Expression levels were normalized to control R/R samples. B , the screening array was filtered for genes significantly induced (≥ 2-fold) in wild-type mice by DEN treatment. Of the 11 genes that were identified, 10 were significantly lower in the homozygous mutant compared with wild-type livers ( n = 3). p21, DR5, WIG, NOXA, and PUMA are referred to as CDKN1A, TNFRSF10B, ZMAT3, PMAIP1, and BBC3, respectively, in (A). C , the mRNA expression levels of 4 prototypical p53-regulated genes were confirmed by independent real-time RT-PCR ( n = 4). Expression levels were normalized relative to wild-type samples. p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); homozygous mutant ( H/H ); and p53 null ( −/− ). Values are mean ± SD. Statistical differences were tested by 1-way ANOVA. * P
    Figure Legend Snippet: Partial loss of p53 transcriptional activity in liver tissue of homozygous mutant p53 334H/H mice treated with DEN. A , color-coded relative expression of 84 p53-regulated genes in untreated (control) and DEN-treated (24 h) mouse livers of the indicated p53 genotype analyzed by RT-PCR screening array ( n = 3). Red and green indicate induced and repressed mRNA levels, respectively. Expression levels were normalized to control R/R samples. B , the screening array was filtered for genes significantly induced (≥ 2-fold) in wild-type mice by DEN treatment. Of the 11 genes that were identified, 10 were significantly lower in the homozygous mutant compared with wild-type livers ( n = 3). p21, DR5, WIG, NOXA, and PUMA are referred to as CDKN1A, TNFRSF10B, ZMAT3, PMAIP1, and BBC3, respectively, in (A). C , the mRNA expression levels of 4 prototypical p53-regulated genes were confirmed by independent real-time RT-PCR ( n = 4). Expression levels were normalized relative to wild-type samples. p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); homozygous mutant ( H/H ); and p53 null ( −/− ). Values are mean ± SD. Statistical differences were tested by 1-way ANOVA. * P

    Techniques Used: Activity Assay, Mutagenesis, Mouse Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

    p53 oligomerization is decreased in p53 334H/H mouse tissues. A , liver tissue lysates were cross-linked with glutaraldehyde, resolved in SDS-PAGE gel, and immunoblotted. Note p53 antibody specificity demonstrated by lack of immunoreactivity in the p53 −/− sample. Protein standards are in kD. Triplicate lanes of each genotype represent liver samples from 3 separate mice. B, fraction of p53 oligomers (T, tetramer; D, dimer; M, monomer) within each lane of immunoblot (A) quantified by densitometry and compared with the respective wild-type oligomer ( n = 3). C , p53 immunoblot of cross-linked liver lysates obtained from mice 6 h after p53 induction by doxorubicin treatment. Duplicate lanes of each genotype represent liver samples from 2 separate mice. D , p53 binding to the p53 response element (p53RE) of p21 in γ-irradiated (γ-IR) mouse liver. ChIP was performed using nonspecific IgG or anti-p53 antibody. p53RE binding is shown relative to wild-type non-specific IgG samples ( n = 3). E, induction of p21 mRNA in the indicated tissues by γ-IR quantified by RT-PCR ( n = 3). Levels were normalized relative to a housekeeping gene TIF . Bone marrow (BM); small intestine. p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); homozygous mutant ( H/H ); and p53 null ( −/− ). Values are mean ± SD. Compared to wild-type samples within each group, statistical differences were tested by 1-way ANOVA. * P
    Figure Legend Snippet: p53 oligomerization is decreased in p53 334H/H mouse tissues. A , liver tissue lysates were cross-linked with glutaraldehyde, resolved in SDS-PAGE gel, and immunoblotted. Note p53 antibody specificity demonstrated by lack of immunoreactivity in the p53 −/− sample. Protein standards are in kD. Triplicate lanes of each genotype represent liver samples from 3 separate mice. B, fraction of p53 oligomers (T, tetramer; D, dimer; M, monomer) within each lane of immunoblot (A) quantified by densitometry and compared with the respective wild-type oligomer ( n = 3). C , p53 immunoblot of cross-linked liver lysates obtained from mice 6 h after p53 induction by doxorubicin treatment. Duplicate lanes of each genotype represent liver samples from 2 separate mice. D , p53 binding to the p53 response element (p53RE) of p21 in γ-irradiated (γ-IR) mouse liver. ChIP was performed using nonspecific IgG or anti-p53 antibody. p53RE binding is shown relative to wild-type non-specific IgG samples ( n = 3). E, induction of p21 mRNA in the indicated tissues by γ-IR quantified by RT-PCR ( n = 3). Levels were normalized relative to a housekeeping gene TIF . Bone marrow (BM); small intestine. p53 R334 genotypes: wild-type ( R/R ); heterozygous mutant ( R/H ); homozygous mutant ( H/H ); and p53 null ( −/− ). Values are mean ± SD. Compared to wild-type samples within each group, statistical differences were tested by 1-way ANOVA. * P

    Techniques Used: SDS Page, Mouse Assay, Binding Assay, Irradiation, Chromatin Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction, Mutagenesis

    21) Product Images from "Three-dimensional perfused human in vitro model of non-alcoholic fatty liver disease"

    Article Title: Three-dimensional perfused human in vitro model of non-alcoholic fatty liver disease

    Journal: World Journal of Gastroenterology

    doi: 10.3748/wjg.v23.i2.204

    Hepatocytes cultured in fat media have altered gene expression profiles. Hepatocytes were cultured in fat or lean conditions for 7 d before total RNA was extracted and gene expression was compared using Fatty Liver RT2 Profiler PCR Arrays. A: Gene expression changes were defined by a fold change > 1.95 and P ≤ 0.05; B: Fold change in expression in fat vs lean condition of key genes, filled bars = fat, white bars = lean. Data are mean ± SD from nine independent cultures (three donors per condition and n = 3 per donor).
    Figure Legend Snippet: Hepatocytes cultured in fat media have altered gene expression profiles. Hepatocytes were cultured in fat or lean conditions for 7 d before total RNA was extracted and gene expression was compared using Fatty Liver RT2 Profiler PCR Arrays. A: Gene expression changes were defined by a fold change > 1.95 and P ≤ 0.05; B: Fold change in expression in fat vs lean condition of key genes, filled bars = fat, white bars = lean. Data are mean ± SD from nine independent cultures (three donors per condition and n = 3 per donor).

    Techniques Used: Cell Culture, Expressing, Polymerase Chain Reaction

    22) Product Images from "Diatom-Derived Polyunsaturated Aldehydes Activate Cell Death in Human Cancer Cell Lines but Not Normal Cells"

    Article Title: Diatom-Derived Polyunsaturated Aldehydes Activate Cell Death in Human Cancer Cell Lines but Not Normal Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0101220

    Effect of the diatom PUAs 2- trans ,4- trans -decadienal (DD), 2- trans ,4- trans -octadienal (OD) and 2- trans ,4- trans -heptadienal (HD) on the human lung adenocarcinoma cell line A549. Control and treated cells double stained with acridine orange and ethidium bromide after 48 µM DD, OD and HD observed at the confocal microscope. Numbers indicate (1) normal cells; (2) early apoptotic cells; (3) late apoptotic cells; (4) necrotic cells (see material and methods for details). Arrows indicate cells with fragmented nuclei.
    Figure Legend Snippet: Effect of the diatom PUAs 2- trans ,4- trans -decadienal (DD), 2- trans ,4- trans -octadienal (OD) and 2- trans ,4- trans -heptadienal (HD) on the human lung adenocarcinoma cell line A549. Control and treated cells double stained with acridine orange and ethidium bromide after 48 µM DD, OD and HD observed at the confocal microscope. Numbers indicate (1) normal cells; (2) early apoptotic cells; (3) late apoptotic cells; (4) necrotic cells (see material and methods for details). Arrows indicate cells with fragmented nuclei.

    Techniques Used: Staining, Microscopy

    23) Product Images from "A transcriptomic approach to elucidate the physiological significance of human cytochrome P450 2S1 in bronchial epithelial cells"

    Article Title: A transcriptomic approach to elucidate the physiological significance of human cytochrome P450 2S1 in bronchial epithelial cells

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-14-833

    Concordance between RNA seq and quantitative PCR array within genes of the mTOR signaling pathway. A subset of 84 genes from the mTOR PCR array is compared between shRNA targets (759 and 984) and methodologies (i.e. RNA-seq and PCR array) using a heat map. Grey indicates no significant change in expression. Gold represents very low expression (i.e. base mean average value is
    Figure Legend Snippet: Concordance between RNA seq and quantitative PCR array within genes of the mTOR signaling pathway. A subset of 84 genes from the mTOR PCR array is compared between shRNA targets (759 and 984) and methodologies (i.e. RNA-seq and PCR array) using a heat map. Grey indicates no significant change in expression. Gold represents very low expression (i.e. base mean average value is

    Techniques Used: RNA Sequencing Assay, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, shRNA, Expressing

    24) Product Images from "Adipocyte arrestin domain-containing 3 protein (Arrdc3) regulates uncoupling protein 1 (Ucp1) expression in white adipose independently of canonical changes in β-adrenergic receptor signaling"

    Article Title: Adipocyte arrestin domain-containing 3 protein (Arrdc3) regulates uncoupling protein 1 (Ucp1) expression in white adipose independently of canonical changes in β-adrenergic receptor signaling

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0173823

    Arrdc3 deletion increases PPAR target gene expression in adipocytes. Quantitative PCR array analysis of gene expression of PPAR target genes, PPAR cofactors and PPARs and associated transcription factors in control versus Arrdc3 SVF-derived adipocytes in vitro (n = 3 mice per group). Only significantly different genes are displayed. *p≤ 0.05.
    Figure Legend Snippet: Arrdc3 deletion increases PPAR target gene expression in adipocytes. Quantitative PCR array analysis of gene expression of PPAR target genes, PPAR cofactors and PPARs and associated transcription factors in control versus Arrdc3 SVF-derived adipocytes in vitro (n = 3 mice per group). Only significantly different genes are displayed. *p≤ 0.05.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Derivative Assay, In Vitro, Mouse Assay

    25) Product Images from "Ischemia/Reperfusion Induces Interferon-Stimulated Gene Expression in Microglia"

    Article Title: Ischemia/Reperfusion Induces Interferon-Stimulated Gene Expression in Microglia

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0725-17.2017

    Microglia upregulate ISGs in response to ischemia/reperfusion in vivo and this response is dependent on IFNAR1 , but not TLR4. WT, TLR4 −/− , and IFNAR1 −/− mice aged 12–14 weeks were subjected to tMCAO/R before brains were collected and processed as described in the Materials and Methods. Microglia were ex vivo sorted by flow cytometry, microglial RNA was isolated, and qRT-PCR was performed with a selected panel of ISG primer/probe sets as described in the Materials and Methods. Expression data were normalized to a panel of housekeeping genes and are presented relative to expression in microglia isolated from contralateral cortex. N = 12–16 mice from four separate ex vivo flow cytometry preparations/sorts for each genotype. # p
    Figure Legend Snippet: Microglia upregulate ISGs in response to ischemia/reperfusion in vivo and this response is dependent on IFNAR1 , but not TLR4. WT, TLR4 −/− , and IFNAR1 −/− mice aged 12–14 weeks were subjected to tMCAO/R before brains were collected and processed as described in the Materials and Methods. Microglia were ex vivo sorted by flow cytometry, microglial RNA was isolated, and qRT-PCR was performed with a selected panel of ISG primer/probe sets as described in the Materials and Methods. Expression data were normalized to a panel of housekeeping genes and are presented relative to expression in microglia isolated from contralateral cortex. N = 12–16 mice from four separate ex vivo flow cytometry preparations/sorts for each genotype. # p

    Techniques Used: In Vivo, Mouse Assay, Ex Vivo, Flow Cytometry, Cytometry, Isolation, Quantitative RT-PCR, Expressing

    26) Product Images from "Molecular mechanism of action of bisphenol and bisphenol A mediated by oestrogen receptor alpha in growth and apoptosis of breast cancer cells"

    Article Title: Molecular mechanism of action of bisphenol and bisphenol A mediated by oestrogen receptor alpha in growth and apoptosis of breast cancer cells

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.12122

    Differential effect of bisphenol (BP) and bisphenol A (BPA) on growth and apoptosis of ERα positive breast cancer cells. (A) Dose-dependent effects of BP, BPA and (oestradiol) E2 on growth of MCF7 cells treated for 6 days as indicated. The black bar denotes the level of DNA in vehicle treated cells over a 6-day period. The growth is measured as amount of DNA present in each well. (* P
    Figure Legend Snippet: Differential effect of bisphenol (BP) and bisphenol A (BPA) on growth and apoptosis of ERα positive breast cancer cells. (A) Dose-dependent effects of BP, BPA and (oestradiol) E2 on growth of MCF7 cells treated for 6 days as indicated. The black bar denotes the level of DNA in vehicle treated cells over a 6-day period. The growth is measured as amount of DNA present in each well. (* P

    Techniques Used:

    27) Product Images from "Systematic Mapping of WNT-FZD Protein Interactions Reveals Functional Selectivity by Distinct WNT-FZD Pairs *"

    Article Title: Systematic Mapping of WNT-FZD Protein Interactions Reveals Functional Selectivity by Distinct WNT-FZD Pairs *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M114.612648

    32D myeloid progenitor cell line expresses little or no endogenous FZDs but does express LRP5/6 co-receptors. A , the bar graph presents the FZD expression profile (FZD 1–10 ) of 32D cells ( hatched gray bars ) and primary microglia cells ( white bars ) determined by qPCR ( n ≥ 3). Error bars represent S.E. GAPDH was used as a housekeeping gene for normalization. By comparing FZD expression in primary microglia cells (previously published by Halleskog et al. )) and 32D cells, it becomes apparent that FZD levels in 32D cells are very low. B , WNT co-receptor profile in 32D cells determined by RT-PCR. LRP5 and LRP6 (but neither RYK nor ROR1/2) are expressed in 32D cells. The negative control consisted of 32D cell cDNA prepared without reverse transcriptase; mouse tail genomic DNA or cDNA from GL261 cells (RYK) was used as a positive control. C , shown are comparative PCR expression profiles of 32D and L929 cells, including WNTs, FZDs, LRPs, DKKs, and SFRPs. The table summarizes the average C t values of two biological replicates. The following genes were not included on the PCR array: Wnt9b , Wnt10b , Fzd9 , Fzd10 , Ryk , Ror1 , and Ror2. D , immunoblotting for HA-tagged FZDs in 32D cells expressing FZD 2 , FZD 4 , or FZD 5 . β-Actin was used as a loading control. The molecular masses of FZDs were determined with Bio-Rad software.
    Figure Legend Snippet: 32D myeloid progenitor cell line expresses little or no endogenous FZDs but does express LRP5/6 co-receptors. A , the bar graph presents the FZD expression profile (FZD 1–10 ) of 32D cells ( hatched gray bars ) and primary microglia cells ( white bars ) determined by qPCR ( n ≥ 3). Error bars represent S.E. GAPDH was used as a housekeeping gene for normalization. By comparing FZD expression in primary microglia cells (previously published by Halleskog et al. )) and 32D cells, it becomes apparent that FZD levels in 32D cells are very low. B , WNT co-receptor profile in 32D cells determined by RT-PCR. LRP5 and LRP6 (but neither RYK nor ROR1/2) are expressed in 32D cells. The negative control consisted of 32D cell cDNA prepared without reverse transcriptase; mouse tail genomic DNA or cDNA from GL261 cells (RYK) was used as a positive control. C , shown are comparative PCR expression profiles of 32D and L929 cells, including WNTs, FZDs, LRPs, DKKs, and SFRPs. The table summarizes the average C t values of two biological replicates. The following genes were not included on the PCR array: Wnt9b , Wnt10b , Fzd9 , Fzd10 , Ryk , Ror1 , and Ror2. D , immunoblotting for HA-tagged FZDs in 32D cells expressing FZD 2 , FZD 4 , or FZD 5 . β-Actin was used as a loading control. The molecular masses of FZDs were determined with Bio-Rad software.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Negative Control, Positive Control, Polymerase Chain Reaction, Software

    28) Product Images from "Doxorubicin induces cardiotoxicity through upregulation of death receptors mediated apoptosis in cardiomyocytes"

    Article Title: Doxorubicin induces cardiotoxicity through upregulation of death receptors mediated apoptosis in cardiomyocytes

    Journal: Scientific Reports

    doi: 10.1038/srep44735

    A working model by which doxorubicin induces cardiotoxicity through upregulation of death receptors mediated apoptosis in cardiomyocytes. Doxorubicin and related anthracyclines appear to be potent inducers of the expression of death receptors (TNFR1, Fas, DR4 and DR5) in cardiomyocytes. The upregulated DRs may undergo clustering or engage their cognate ligands, thereby triggering a caspase cascade and ultimate apoptosis in cardiomyocytes. The elevated serum levels of specific TNF cytokines (e.g., TRAIL), which could occur under certain disease and treatment conditions, may be predictive of the risk of cardiotoxicity in individual patients prior to the administration of doxorubicin or anthracycline agents.
    Figure Legend Snippet: A working model by which doxorubicin induces cardiotoxicity through upregulation of death receptors mediated apoptosis in cardiomyocytes. Doxorubicin and related anthracyclines appear to be potent inducers of the expression of death receptors (TNFR1, Fas, DR4 and DR5) in cardiomyocytes. The upregulated DRs may undergo clustering or engage their cognate ligands, thereby triggering a caspase cascade and ultimate apoptosis in cardiomyocytes. The elevated serum levels of specific TNF cytokines (e.g., TRAIL), which could occur under certain disease and treatment conditions, may be predictive of the risk of cardiotoxicity in individual patients prior to the administration of doxorubicin or anthracycline agents.

    Techniques Used: Expressing

    29) Product Images from "An Autocrine Cytokine/JAK/STAT-Signaling Induces Kynurenine Synthesis in Multidrug Resistant Human Cancer Cells"

    Article Title: An Autocrine Cytokine/JAK/STAT-Signaling Induces Kynurenine Synthesis in Multidrug Resistant Human Cancer Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0126159

    Multidrug resistant cells have a higher activity of JAK/STAT signaling than chemosensitive cells. A. The cDNA from A549 and A549/dx cells was analyzed by a PCR array specific for JAK/STAT signaling, as reported under Materials and methods. The fold regulation of the 83 genes analyzed, expressed in logarithmic scale, is represented in a colorimetric scale. The figure is the mean of 4 experiments. B. The cells were lysed and subjected to the Western blot analysis for phospho(Tyr 1022/1023)-JAK1, JAK1, phospho(Tyr701)-STAT1, STAT1, phospho(Tyr705)-STAT3, STAT3. The β-tubulin expression was used as control of equal protein loading. The figure is representative of 3 experiments with similar results. C. The expression of PIAS1, PIAS3, phospho(Tyr701)-STAT1, STAT1, phospho(Tyr705)-STAT3, STAT3 in nuclear extracts was measured by Western blotting. The TBP expression was used as control of equal protein loading. The figure is representative of 3 experiments with similar results.
    Figure Legend Snippet: Multidrug resistant cells have a higher activity of JAK/STAT signaling than chemosensitive cells. A. The cDNA from A549 and A549/dx cells was analyzed by a PCR array specific for JAK/STAT signaling, as reported under Materials and methods. The fold regulation of the 83 genes analyzed, expressed in logarithmic scale, is represented in a colorimetric scale. The figure is the mean of 4 experiments. B. The cells were lysed and subjected to the Western blot analysis for phospho(Tyr 1022/1023)-JAK1, JAK1, phospho(Tyr701)-STAT1, STAT1, phospho(Tyr705)-STAT3, STAT3. The β-tubulin expression was used as control of equal protein loading. The figure is representative of 3 experiments with similar results. C. The expression of PIAS1, PIAS3, phospho(Tyr701)-STAT1, STAT1, phospho(Tyr705)-STAT3, STAT3 in nuclear extracts was measured by Western blotting. The TBP expression was used as control of equal protein loading. The figure is representative of 3 experiments with similar results.

    Techniques Used: Activity Assay, Polymerase Chain Reaction, Western Blot, Expressing

    30) Product Images from "Delayed triggering of oestrogen induced apoptosis that contrasts with rapid paclitaxel-induced breast cancer cell death"

    Article Title: Delayed triggering of oestrogen induced apoptosis that contrasts with rapid paclitaxel-induced breast cancer cell death

    Journal: British Journal of Cancer

    doi: 10.1038/bjc.2014.50

    Differential apoptotic effects of E 2 and paclitaxel. MCF7:5C cells were treated with control or ( A ) paclitaxel (1 μ M ) for 12 and 24 h or ( B ) E 2 (1 n M ) for 72 h, and then stained with annexin V-FITC and PI and analysed by flow cytometry. Viable cells (left lower quadrant) are annexin V-FITC− and PI−, early apoptotic cells (right lower quadrant) are annexin V-FITC+ and PI−, dead cells (left upper quadrant) are PI+ and late apoptotic cells (right upper quadrant) are annexin V-FITC+ and PI+. Increased staining for apoptosis is observed maximally in the right upper quadrant.
    Figure Legend Snippet: Differential apoptotic effects of E 2 and paclitaxel. MCF7:5C cells were treated with control or ( A ) paclitaxel (1 μ M ) for 12 and 24 h or ( B ) E 2 (1 n M ) for 72 h, and then stained with annexin V-FITC and PI and analysed by flow cytometry. Viable cells (left lower quadrant) are annexin V-FITC− and PI−, early apoptotic cells (right lower quadrant) are annexin V-FITC+ and PI−, dead cells (left upper quadrant) are PI+ and late apoptotic cells (right upper quadrant) are annexin V-FITC+ and PI+. Increased staining for apoptosis is observed maximally in the right upper quadrant.

    Techniques Used: Staining, Flow Cytometry, Cytometry

    Deciphering the trigger point for E 2 -induced apoptosis. Cells were treated with vehicle (Veh) or E 2 (1 n M ) alone, and 1 μ M 4OHT was added and used to block and reverse E 2 action at 6, 12, 24, 36, 48, 60, 72, 84 and 96 h. The cells were harvested after 7 days of treatment. The extent of apoptosis was determined by measuring the DNA content of the remaining cells in each well. The experiment was done in triplicates, and the data represent the mean of three independent experiments with 95% confidence intervals. The trigger point for E 2 -mediated apoptosis was elucidated at the time when the apoptotic effects of E 2 could not be blocked by 4OHT.
    Figure Legend Snippet: Deciphering the trigger point for E 2 -induced apoptosis. Cells were treated with vehicle (Veh) or E 2 (1 n M ) alone, and 1 μ M 4OHT was added and used to block and reverse E 2 action at 6, 12, 24, 36, 48, 60, 72, 84 and 96 h. The cells were harvested after 7 days of treatment. The extent of apoptosis was determined by measuring the DNA content of the remaining cells in each well. The experiment was done in triplicates, and the data represent the mean of three independent experiments with 95% confidence intervals. The trigger point for E 2 -mediated apoptosis was elucidated at the time when the apoptotic effects of E 2 could not be blocked by 4OHT.

    Techniques Used: Blocking Assay

    E 2 activates both mitochondrial and extrinsic pathway of apoptosis, whereas paclitaxel activates only the extrinsic pathway. MCF7:5C cells were treated with vehicle (Veh), 1 n M E2, 1 μ M 4OHT or combination treatment of E 2 and 4OHT for 24, 36 and 48 h. Total RNA was reverse transcribed and assessed for ( A ) BIM and ( B ) TNF gene expression. Induction of ( C ) BIM and ( D ) TNF mRNA was determined in MCF7:5C cells treated with either Veh or 1 μ M paclitaxel for 12 and 24 h using RT–PCR. PCR data values are presented as fold difference versus Veh-treated cells±s.e.m. (** P
    Figure Legend Snippet: E 2 activates both mitochondrial and extrinsic pathway of apoptosis, whereas paclitaxel activates only the extrinsic pathway. MCF7:5C cells were treated with vehicle (Veh), 1 n M E2, 1 μ M 4OHT or combination treatment of E 2 and 4OHT for 24, 36 and 48 h. Total RNA was reverse transcribed and assessed for ( A ) BIM and ( B ) TNF gene expression. Induction of ( C ) BIM and ( D ) TNF mRNA was determined in MCF7:5C cells treated with either Veh or 1 μ M paclitaxel for 12 and 24 h using RT–PCR. PCR data values are presented as fold difference versus Veh-treated cells±s.e.m. (** P

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

    31) Product Images from "Innate And Adaptive Immunity are Progressively Activated in Parallel with Renal Injury in the 5/6 Renal Ablation Model"

    Article Title: Innate And Adaptive Immunity are Progressively Activated in Parallel with Renal Injury in the 5/6 Renal Ablation Model

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-02915-6

    Quantitative PCR analysis (qPCR) for Tlr4 ( A ), Nlrp3 ( B ), Casp1 ( C ) and Il1b ( D ) gene expression. Results are presented as median plus range. The number of analyzed animals is represented next to the name of each group. Differences among groups were analyzed by one-way ANOVA with Newman-Keuls post test. a p
    Figure Legend Snippet: Quantitative PCR analysis (qPCR) for Tlr4 ( A ), Nlrp3 ( B ), Casp1 ( C ) and Il1b ( D ) gene expression. Results are presented as median plus range. The number of analyzed animals is represented next to the name of each group. Differences among groups were analyzed by one-way ANOVA with Newman-Keuls post test. a p

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing

    32) Product Images from "The oncogenic role of the In1-ghrelin splicing variant in prostate cancer aggressiveness"

    Article Title: The oncogenic role of the In1-ghrelin splicing variant in prostate cancer aggressiveness

    Journal: Molecular Cancer

    doi: 10.1186/s12943-017-0713-9

    Gene expression effects of ghrelin and In1-ghrelin overexpression in PC-3 and derived xenografted tumors. a . Results from the RT2 Prostate-Cancer PCR Array which evaluates the expression of 84 genes involved in prostate cancer development and progression performed in ghrelin and In1-ghrelin-stably transfected PC-3-cells compared with control-mock PC-3-cells. The graphs indicate those genes which expression change ≥ 1.5-fold; b . Validation by qPCR of genes dysregulated in the RT2 Prostate-Cancer PCR Array using different cell preparations ( n ≥ 3) and new sets of primers; c . Validation by Western blot of the changes observed in the previous analysis; d . Expression of angiogenic factors in In1-ghrelin-stably transfected PC-3-cells and native ghrelin-stably transfected PC-3 cells compared with control-mock PC-3-cells.; e . Expression of angiogenic factors in xenografted tumors of stably transfected-PC-3 cells. f . CAV1, LOXL1, SFRP1 and IL-6 mRNA expression levels in mock, ghrelin and In1-ghrelin transfected PC-3-derived xenografted tumors. Results were normalized with ACTB. All preparations were repeated at least three times ( n ≥ 3). (* p
    Figure Legend Snippet: Gene expression effects of ghrelin and In1-ghrelin overexpression in PC-3 and derived xenografted tumors. a . Results from the RT2 Prostate-Cancer PCR Array which evaluates the expression of 84 genes involved in prostate cancer development and progression performed in ghrelin and In1-ghrelin-stably transfected PC-3-cells compared with control-mock PC-3-cells. The graphs indicate those genes which expression change ≥ 1.5-fold; b . Validation by qPCR of genes dysregulated in the RT2 Prostate-Cancer PCR Array using different cell preparations ( n ≥ 3) and new sets of primers; c . Validation by Western blot of the changes observed in the previous analysis; d . Expression of angiogenic factors in In1-ghrelin-stably transfected PC-3-cells and native ghrelin-stably transfected PC-3 cells compared with control-mock PC-3-cells.; e . Expression of angiogenic factors in xenografted tumors of stably transfected-PC-3 cells. f . CAV1, LOXL1, SFRP1 and IL-6 mRNA expression levels in mock, ghrelin and In1-ghrelin transfected PC-3-derived xenografted tumors. Results were normalized with ACTB. All preparations were repeated at least three times ( n ≥ 3). (* p

    Techniques Used: Expressing, Over Expression, Derivative Assay, Polymerase Chain Reaction, Stable Transfection, Transfection, Real-time Polymerase Chain Reaction, Western Blot

    Expression of ghrelin and In1-ghrelin in prostate cancer. a . Ghrelin or In1-ghrelin mRNA expression in biopsies from patients with high-risk PCa ( n = 52) and normal prostate from patients that underwent cystoprostatectomy ( n = 12). Expression levels were determined by qPCR and adjusted by a normalization factor (NF) calculated from ACTB and GAPDH expression levels; b . ROC curve analysis to determine the accuracy of ghrelin or In1-ghrelin mRNA expression as diagnostic test to discriminate between high-risk PCa patients and controls using the same cohort. c . Correlations between ghrelin or In1-ghrelin expression with Ki-67 mRNA expression in PCa patients d . Correlations between ghrelin or In1-ghrelin expression with GOAT enzyme mRNA expression in PCa patients. e . Correlations between ghrelin or In1-ghrelin with PSA in PCa patients; f . Expression of acylated-ghrelin (ELISA) or acylated In1-ghrelin (RIA) in the plasma of patients with PCa ( n = 30) and controls ( n = 20); g . ROC curve analysis to determine the accuracy of acylated ghrelin or acylated In1-ghrelin plasmatic protein expression as diagnostic test to discriminate between PCa patients and controls. h . Ghrelin mRNA expression levels in normal-like prostate cell line (RWPE-1) or PCa cell lines; i . In1-ghrelin mRNA expression levels in normal-like prostate cell line (RWPE-1) or PCa cell lines. Absolute mRNA levels from different passages ( n ≥ 3) were determined by qPCR and adjusted by ACTB. Asterisks (* p
    Figure Legend Snippet: Expression of ghrelin and In1-ghrelin in prostate cancer. a . Ghrelin or In1-ghrelin mRNA expression in biopsies from patients with high-risk PCa ( n = 52) and normal prostate from patients that underwent cystoprostatectomy ( n = 12). Expression levels were determined by qPCR and adjusted by a normalization factor (NF) calculated from ACTB and GAPDH expression levels; b . ROC curve analysis to determine the accuracy of ghrelin or In1-ghrelin mRNA expression as diagnostic test to discriminate between high-risk PCa patients and controls using the same cohort. c . Correlations between ghrelin or In1-ghrelin expression with Ki-67 mRNA expression in PCa patients d . Correlations between ghrelin or In1-ghrelin expression with GOAT enzyme mRNA expression in PCa patients. e . Correlations between ghrelin or In1-ghrelin with PSA in PCa patients; f . Expression of acylated-ghrelin (ELISA) or acylated In1-ghrelin (RIA) in the plasma of patients with PCa ( n = 30) and controls ( n = 20); g . ROC curve analysis to determine the accuracy of acylated ghrelin or acylated In1-ghrelin plasmatic protein expression as diagnostic test to discriminate between PCa patients and controls. h . Ghrelin mRNA expression levels in normal-like prostate cell line (RWPE-1) or PCa cell lines; i . In1-ghrelin mRNA expression levels in normal-like prostate cell line (RWPE-1) or PCa cell lines. Absolute mRNA levels from different passages ( n ≥ 3) were determined by qPCR and adjusted by ACTB. Asterisks (* p

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

    33) Product Images from "The oncogenic role of the In1-ghrelin splicing variant in prostate cancer aggressiveness"

    Article Title: The oncogenic role of the In1-ghrelin splicing variant in prostate cancer aggressiveness

    Journal: Molecular Cancer

    doi: 10.1186/s12943-017-0713-9

    Expression of ghrelin and In1-ghrelin in prostate cancer. a . Ghrelin or In1-ghrelin mRNA expression in biopsies from patients with high-risk PCa ( n = 52) and normal prostate from patients that underwent cystoprostatectomy ( n = 12). Expression levels were determined by qPCR and adjusted by a normalization factor (NF) calculated from ACTB and GAPDH expression levels; b . ROC curve analysis to determine the accuracy of ghrelin or In1-ghrelin mRNA expression as diagnostic test to discriminate between high-risk PCa patients and controls using the same cohort. c . Correlations between ghrelin or In1-ghrelin expression with Ki-67 mRNA expression in PCa patients d . Correlations between ghrelin or In1-ghrelin expression with GOAT enzyme mRNA expression in PCa patients. e . Correlations between ghrelin or In1-ghrelin with PSA in PCa patients; f . Expression of acylated-ghrelin (ELISA) or acylated In1-ghrelin (RIA) in the plasma of patients with PCa ( n = 30) and controls ( n = 20); g . ROC curve analysis to determine the accuracy of acylated ghrelin or acylated In1-ghrelin plasmatic protein expression as diagnostic test to discriminate between PCa patients and controls. h . Ghrelin mRNA expression levels in normal-like prostate cell line (RWPE-1) or PCa cell lines; i . In1-ghrelin mRNA expression levels in normal-like prostate cell line (RWPE-1) or PCa cell lines. Absolute mRNA levels from different passages ( n ≥ 3) were determined by qPCR and adjusted by ACTB. Asterisks (* p
    Figure Legend Snippet: Expression of ghrelin and In1-ghrelin in prostate cancer. a . Ghrelin or In1-ghrelin mRNA expression in biopsies from patients with high-risk PCa ( n = 52) and normal prostate from patients that underwent cystoprostatectomy ( n = 12). Expression levels were determined by qPCR and adjusted by a normalization factor (NF) calculated from ACTB and GAPDH expression levels; b . ROC curve analysis to determine the accuracy of ghrelin or In1-ghrelin mRNA expression as diagnostic test to discriminate between high-risk PCa patients and controls using the same cohort. c . Correlations between ghrelin or In1-ghrelin expression with Ki-67 mRNA expression in PCa patients d . Correlations between ghrelin or In1-ghrelin expression with GOAT enzyme mRNA expression in PCa patients. e . Correlations between ghrelin or In1-ghrelin with PSA in PCa patients; f . Expression of acylated-ghrelin (ELISA) or acylated In1-ghrelin (RIA) in the plasma of patients with PCa ( n = 30) and controls ( n = 20); g . ROC curve analysis to determine the accuracy of acylated ghrelin or acylated In1-ghrelin plasmatic protein expression as diagnostic test to discriminate between PCa patients and controls. h . Ghrelin mRNA expression levels in normal-like prostate cell line (RWPE-1) or PCa cell lines; i . In1-ghrelin mRNA expression levels in normal-like prostate cell line (RWPE-1) or PCa cell lines. Absolute mRNA levels from different passages ( n ≥ 3) were determined by qPCR and adjusted by ACTB. Asterisks (* p

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

    34) Product Images from "Transient PP2A inhibition alleviates normal tissue stem cell susceptibility to cell death during radiotherapy"

    Article Title: Transient PP2A inhibition alleviates normal tissue stem cell susceptibility to cell death during radiotherapy

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-0559-0

    PP2A inhibition decreases radiosensitivity of stem cells in ex vivo organoid cultures. a Shown are optical microscope images of murine intestinal organoids (upper panel) and murine neurospheres (lower panel) untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Pictures have been taken 48 h after IR treatment. b Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl) or PP2A siRNA (siPP2A). Percentage of cells positive for stem cell marker SSEA1 and apoptotic marker CC-3 was calculated from total SSEA1 positive cells. Scoring was performed 4 h after irradiation. c Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Percentage of cells Lgr5-GFP positive was detected by flow cytometry. Scoring was performed 4 h after irradiation. d Murine hematopoietic stem cells were treated with Cal A or left untreated. Cells were irradiated with 2 Gy and apoptosis analyzed after 16 h by Annexin V labeling. e Human neuroprogenitors were treated with Cal A or left untreated. Cells were irradiated with 6 Gy and apoptosis analyzed after 16 h by Annexin V labeling. Scale bar = 100 μm. Error bars indicate SD; * p
    Figure Legend Snippet: PP2A inhibition decreases radiosensitivity of stem cells in ex vivo organoid cultures. a Shown are optical microscope images of murine intestinal organoids (upper panel) and murine neurospheres (lower panel) untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Pictures have been taken 48 h after IR treatment. b Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl) or PP2A siRNA (siPP2A). Percentage of cells positive for stem cell marker SSEA1 and apoptotic marker CC-3 was calculated from total SSEA1 positive cells. Scoring was performed 4 h after irradiation. c Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Percentage of cells Lgr5-GFP positive was detected by flow cytometry. Scoring was performed 4 h after irradiation. d Murine hematopoietic stem cells were treated with Cal A or left untreated. Cells were irradiated with 2 Gy and apoptosis analyzed after 16 h by Annexin V labeling. e Human neuroprogenitors were treated with Cal A or left untreated. Cells were irradiated with 6 Gy and apoptosis analyzed after 16 h by Annexin V labeling. Scale bar = 100 μm. Error bars indicate SD; * p

    Techniques Used: Inhibition, Ex Vivo, Microscopy, Irradiation, Marker, Flow Cytometry, Cytometry, Labeling

    35) Product Images from "Insights into the Staphylococcus aureus-Host Interface: Global Changes in Host and Pathogen Gene Expression in a Rabbit Skin Infection Model"

    Article Title: Insights into the Staphylococcus aureus-Host Interface: Global Changes in Host and Pathogen Gene Expression in a Rabbit Skin Infection Model

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0117713

    S . aureus skin infection causes changes in transcripts encoding molecules involved in Th lymphocyte differentiation. The networks/functional analyses were generated through the use of QIAGEN’s Ingenuity Pathway Analysis (IPA, QIAGEN Redwood City, www.qiagen.com/ingenuity ). The heat map representing changes in transcript levels of molecules on day 1, 3, 6, 10 and 14 during 14-day course of infection, was created using Adobe Illustrator CS5.1 software. Empty symbols indicate molecules not evaluated by the RT² Profiler PCR Array.
    Figure Legend Snippet: S . aureus skin infection causes changes in transcripts encoding molecules involved in Th lymphocyte differentiation. The networks/functional analyses were generated through the use of QIAGEN’s Ingenuity Pathway Analysis (IPA, QIAGEN Redwood City, www.qiagen.com/ingenuity ). The heat map representing changes in transcript levels of molecules on day 1, 3, 6, 10 and 14 during 14-day course of infection, was created using Adobe Illustrator CS5.1 software. Empty symbols indicate molecules not evaluated by the RT² Profiler PCR Array.

    Techniques Used: Infection, Functional Assay, Generated, Indirect Immunoperoxidase Assay, Software, Polymerase Chain Reaction

    36) Product Images from "Extremely low frequency pulsed electromagnetic fields cause antioxidative defense mechanisms in human osteoblasts via induction of •O2− and H2O2"

    Article Title: Extremely low frequency pulsed electromagnetic fields cause antioxidative defense mechanisms in human osteoblasts via induction of •O2− and H2O2

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-14983-9

    ELF-PEMF exposure induces gene expression of intracellular antioxidative enzymes in hOBs. ( a ) Basal expression levels of oxidative stress (•O 2 − and H 2 O 2 ) related genes in hOBs was determined using the RT² Profiler PCR Array human oxidative stress plus (Qiagen, Hilden, Germany). ( b – e ) Semi-quantitative RT-PCR revealed that expression of SOD2, CAT, GPX3 and GSR was increased by repetitive exposures to ELF-PEMF (7 min per day) during the osteogenic differentiation of hOBs. Densitometric analysis (ImageJ software) was performed with individual samples (N = 12) analyzed twice (n = 2) to reduce small loading differences. The mean signal intensity of cells on day 0 was set as reference. * p
    Figure Legend Snippet: ELF-PEMF exposure induces gene expression of intracellular antioxidative enzymes in hOBs. ( a ) Basal expression levels of oxidative stress (•O 2 − and H 2 O 2 ) related genes in hOBs was determined using the RT² Profiler PCR Array human oxidative stress plus (Qiagen, Hilden, Germany). ( b – e ) Semi-quantitative RT-PCR revealed that expression of SOD2, CAT, GPX3 and GSR was increased by repetitive exposures to ELF-PEMF (7 min per day) during the osteogenic differentiation of hOBs. Densitometric analysis (ImageJ software) was performed with individual samples (N = 12) analyzed twice (n = 2) to reduce small loading differences. The mean signal intensity of cells on day 0 was set as reference. * p

    Techniques Used: Expressing, Polymerase Chain Reaction, Quantitative RT-PCR, Software

    37) Product Images from "A transcriptomic approach to elucidate the physiological significance of human cytochrome P450 2S1 in bronchial epithelial cells"

    Article Title: A transcriptomic approach to elucidate the physiological significance of human cytochrome P450 2S1 in bronchial epithelial cells

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-14-833

    Concordance between RNA seq and quantitative PCR array within genes of the mTOR signaling pathway. A subset of 84 genes from the mTOR PCR array is compared between shRNA targets (759 and 984) and methodologies (i.e. RNA-seq and PCR array) using a heat map. Grey indicates no significant change in expression. Gold represents very low expression (i.e. base mean average value is
    Figure Legend Snippet: Concordance between RNA seq and quantitative PCR array within genes of the mTOR signaling pathway. A subset of 84 genes from the mTOR PCR array is compared between shRNA targets (759 and 984) and methodologies (i.e. RNA-seq and PCR array) using a heat map. Grey indicates no significant change in expression. Gold represents very low expression (i.e. base mean average value is

    Techniques Used: RNA Sequencing Assay, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, shRNA, Expressing

    38) Product Images from "CHAC2, downregulated in gastric and colorectal cancers, acted as a tumor suppressor inducing apoptosis and autophagy through unfolded protein response"

    Article Title: CHAC2, downregulated in gastric and colorectal cancers, acted as a tumor suppressor inducing apoptosis and autophagy through unfolded protein response

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2017.405

    Analysis of CHAC2 expression and its degradation pathway in gastric and colorectal cancer cell lines. ( a ) The expression of CHAC2 in gastric and colorectal cancer cell lines was determined by RT-PCR and western bolt, using GAPDH as a control. ( b ) CHAC2 expression in stably transfected cells confirmed by RT-PCR and western blot. ( c ) Evaluation of CHAC2 expression after treatment of 5-FU or BFA. ( d ) Analysis of the potential degradation pathway of CHAC2 using different inhibitors. AGS-CHAC2 and SW620-CHAC2 cells were treated with 20 μ g/ml CHX with or without MG-132 (20 μ M) or Leu (100 μ M) or CQ (50 mM) in a time course. Cell lysates were subjected to western blot with primary antibodies including CHAC2, p-ERK and GAPDH. ( e ) The effect of ubiquitin on CHAC2 expression. AGS-CHAC2 and SW620-CHAC2 cells were incubated with indicated rhHA-ubiquitin for 12 h, and cell lysates were subjected to western blot with primary antibodies including CHAC2, HA tag andGAPDH. ( f ) 12 maximal upregulated genes checked by the Human Ubiquitination Pathway RT 2 Profiler PCR Array were shown in the bar graph. Among these genes, RNF148 expression significantly increased nearly 1863 times in SW620 as compared to SW48. ( g ) The expression of RNF148 in gastric and colorectal cancer cell lines was determined by RT-PCR. ( h ) Evaluation of RNF148 expression after treatment of 5-FU or BFA. 5-FU, 5-fluorouracil; BFA, Brefeldin A; CHX, Cycloheximide; Leu, leupeptin; CQ, Chloroquine; Ubi, rhHA-ubiquitin
    Figure Legend Snippet: Analysis of CHAC2 expression and its degradation pathway in gastric and colorectal cancer cell lines. ( a ) The expression of CHAC2 in gastric and colorectal cancer cell lines was determined by RT-PCR and western bolt, using GAPDH as a control. ( b ) CHAC2 expression in stably transfected cells confirmed by RT-PCR and western blot. ( c ) Evaluation of CHAC2 expression after treatment of 5-FU or BFA. ( d ) Analysis of the potential degradation pathway of CHAC2 using different inhibitors. AGS-CHAC2 and SW620-CHAC2 cells were treated with 20 μ g/ml CHX with or without MG-132 (20 μ M) or Leu (100 μ M) or CQ (50 mM) in a time course. Cell lysates were subjected to western blot with primary antibodies including CHAC2, p-ERK and GAPDH. ( e ) The effect of ubiquitin on CHAC2 expression. AGS-CHAC2 and SW620-CHAC2 cells were incubated with indicated rhHA-ubiquitin for 12 h, and cell lysates were subjected to western blot with primary antibodies including CHAC2, HA tag andGAPDH. ( f ) 12 maximal upregulated genes checked by the Human Ubiquitination Pathway RT 2 Profiler PCR Array were shown in the bar graph. Among these genes, RNF148 expression significantly increased nearly 1863 times in SW620 as compared to SW48. ( g ) The expression of RNF148 in gastric and colorectal cancer cell lines was determined by RT-PCR. ( h ) Evaluation of RNF148 expression after treatment of 5-FU or BFA. 5-FU, 5-fluorouracil; BFA, Brefeldin A; CHX, Cycloheximide; Leu, leupeptin; CQ, Chloroquine; Ubi, rhHA-ubiquitin

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Stable Transfection, Transfection, Incubation, Polymerase Chain Reaction

    39) Product Images from "Ischemia/Reperfusion Induces Interferon-Stimulated Gene Expression in Microglia"

    Article Title: Ischemia/Reperfusion Induces Interferon-Stimulated Gene Expression in Microglia

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0725-17.2017

    Microglia upregulate ISGs in response to ischemia/reperfusion in vivo and this response is dependent on IFNAR1 , but not TLR4. WT, TLR4 −/− , and IFNAR1 −/− mice aged 12–14 weeks were subjected to tMCAO/R before brains were collected and processed as described in the Materials and Methods. Microglia were ex vivo sorted by flow cytometry, microglial RNA was isolated, and qRT-PCR was performed with a selected panel of ISG primer/probe sets as described in the Materials and Methods. Expression data were normalized to a panel of housekeeping genes and are presented relative to expression in microglia isolated from contralateral cortex. N = 12–16 mice from four separate ex vivo flow cytometry preparations/sorts for each genotype. # p
    Figure Legend Snippet: Microglia upregulate ISGs in response to ischemia/reperfusion in vivo and this response is dependent on IFNAR1 , but not TLR4. WT, TLR4 −/− , and IFNAR1 −/− mice aged 12–14 weeks were subjected to tMCAO/R before brains were collected and processed as described in the Materials and Methods. Microglia were ex vivo sorted by flow cytometry, microglial RNA was isolated, and qRT-PCR was performed with a selected panel of ISG primer/probe sets as described in the Materials and Methods. Expression data were normalized to a panel of housekeeping genes and are presented relative to expression in microglia isolated from contralateral cortex. N = 12–16 mice from four separate ex vivo flow cytometry preparations/sorts for each genotype. # p

    Techniques Used: In Vivo, Mouse Assay, Ex Vivo, Flow Cytometry, Cytometry, Isolation, Quantitative RT-PCR, Expressing

    40) Product Images from "Preconditioning by Hydrogen Peroxide Enhances Multiple Properties of Human Decidua Basalis Mesenchymal Stem/Multipotent Stromal Cells"

    Article Title: Preconditioning by Hydrogen Peroxide Enhances Multiple Properties of Human Decidua Basalis Mesenchymal Stem/Multipotent Stromal Cells

    Journal: Stem Cells International

    doi: 10.1155/2018/6480793

    List of stress-related genes modulated in DBMSCs after exposure to H2 O2 .
    Figure Legend Snippet: List of stress-related genes modulated in DBMSCs after exposure to H2 O2 .

    Techniques Used:

    Related Articles

    Polymerase Chain Reaction:

    Article Title: Adipocyte arrestin domain-containing 3 protein (Arrdc3) regulates uncoupling protein 1 (Ucp1) expression in white adipose independently of canonical changes in β-adrenergic receptor signaling
    Article Snippet: .. PPAR target gene expression was performed using RT2 Profiler PCR Array Mouse PPAR Targets (Qiagen cat. no. 330231 PAMM-149ZA). .. Western analysis Tissues were homogenized in RIPA buffer (150 mM NaCl, 0.1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 50 mM Tris-HCl pH 8.0) supplemented with protease inhibitor (cOmplete Mini, Roche).

    Article Title: Rationalized inhibition of mixed lineage kinase 3 and CD70 enhances life span and antitumor efficacy of CD8+ T cells
    Article Snippet: .. The cDNA was used for RT² Profiler PCR array mouse apoptosis (Qiagen) using real-time PCR (ABI StepOnePlus). .. The RT² Profiler PCR array data were analyzed by using SA Biosciences software.

    Article Title: Transient PP2A inhibition alleviates normal tissue stem cell susceptibility to cell death during radiotherapy
    Article Snippet: .. To obtain a more holistic view of PP2Ai effects on apoptotic network proteins, we performed RT-PCR analysis on a panel of apoptosis regulatory genes (RT² Profiler™ PCR Array Mouse Apoptosis from Qiagen). .. This analysis showed a clear reduction in expression of several genes involved in the apoptotic pathway in ES treated with Cal A compared to the 6 Gy control, with a dramatic decrease in caspase-4 and Fas expression (Fig. ).

    Article Title: Expression of p53 Target Genes in the Early Phase of Long-Term Potentiation in the Rat Hippocampal CA1 Area
    Article Snippet: .. 2.3. cDNA Synthesis and Real-Time PCR To identify the participation of p53 in transcriptional regulation during the early phase of LTP, a Rat p53 Signaling Pathway RT² Profiler PCR Array (Qiagen) was used. .. Total RNA was isolated using an AllPrep DNA/RNA/Protein Mini Kit (Qiagen) according to manufacturer's protocol.

    Article Title: CHAC2, downregulated in gastric and colorectal cancers, acted as a tumor suppressor inducing apoptosis and autophagy through unfolded protein response
    Article Snippet: .. The human ubiquitination pathway RT2 profiler PCR array The Human Ubiquitination Pathway RT2 Profiler PCR Array (Cat. no. 330231, Qiagen, Hilden, German) was used to profile the expression of 84 key genes involved in the regulated degradation of cellular proteins by the ubiquitin-proteasome. ..

    Article Title: GADD45A and CDKN1A are involved in apoptosis and cell cycle modulatory effects of viscumTT with further inactivation of the STAT3 pathway
    Article Snippet: .. RT² Profiler™ PCR Array of cell cycle genes Alterations in 84 cell cycle genes were analyzed by RT² Profiler™ PCR Array Human Cell Cycle (Qiagen, Hilden, Germany). ..

    Article Title: Thyroid hormone induces progression and invasiveness of squamous cell carcinomas by promoting a ZEB-1/E-cadherin switch
    Article Snippet: .. We profiled the expression of 84 genes whose expression is related to the EMT using the human EMT RT² Profiler™ PCR Array (Qiagen). .. Interestingly, most of the EMT genes were modified by D3-depletion (Fig. ).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Molecular mechanism of action of bisphenol and bisphenol A mediated by oestrogen receptor alpha in growth and apoptosis of breast cancer cells
    Article Snippet: .. RT-PCR profiler assay kits for apoptosis was used from a commercial vendor which uses 384 well plates to profile the expression of 370 apoptosis related human genes (Qiagen; SABiosciences Corp, Fredrick, MD, USA; Cat#330231 PAHS-3012E). .. Briefly, MCF7 : 5C cells were treated with E2 (10−9 M) for 24, 48 and 72 h or with indicated compounds (in triplicate) for 48 h and total RNA was isolated using the method mentioned earlier.

    Article Title: Transient PP2A inhibition alleviates normal tissue stem cell susceptibility to cell death during radiotherapy
    Article Snippet: .. To obtain a more holistic view of PP2Ai effects on apoptotic network proteins, we performed RT-PCR analysis on a panel of apoptosis regulatory genes (RT² Profiler™ PCR Array Mouse Apoptosis from Qiagen). .. This analysis showed a clear reduction in expression of several genes involved in the apoptotic pathway in ES treated with Cal A compared to the 6 Gy control, with a dramatic decrease in caspase-4 and Fas expression (Fig. ).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Molecular mechanism of action of bisphenol and bisphenol A mediated by oestrogen receptor alpha in growth and apoptosis of breast cancer cells
    Article Snippet: .. RT-PCR profiler assay kits for apoptosis was used from a commercial vendor which uses 384 well plates to profile the expression of 370 apoptosis related human genes (Qiagen; SABiosciences Corp, Fredrick, MD, USA; Cat330231 PAHS-3012E). .. Briefly, MCF7 : 5C cells were treated with E2 (10−9 M) for 24, 48 and 72 h or with indicated compounds (in triplicate) for 48 h and total RNA was isolated using the method mentioned earlier.

    Real-time Polymerase Chain Reaction:

    Article Title: Rationalized inhibition of mixed lineage kinase 3 and CD70 enhances life span and antitumor efficacy of CD8+ T cells
    Article Snippet: .. The cDNA was used for RT² Profiler PCR array mouse apoptosis (Qiagen) using real-time PCR (ABI StepOnePlus). .. The RT² Profiler PCR array data were analyzed by using SA Biosciences software.

    Article Title: Expression of p53 Target Genes in the Early Phase of Long-Term Potentiation in the Rat Hippocampal CA1 Area
    Article Snippet: .. 2.3. cDNA Synthesis and Real-Time PCR To identify the participation of p53 in transcriptional regulation during the early phase of LTP, a Rat p53 Signaling Pathway RT² Profiler PCR Array (Qiagen) was used. .. Total RNA was isolated using an AllPrep DNA/RNA/Protein Mini Kit (Qiagen) according to manufacturer's protocol.

    Expressing:

    Article Title: Adipocyte arrestin domain-containing 3 protein (Arrdc3) regulates uncoupling protein 1 (Ucp1) expression in white adipose independently of canonical changes in β-adrenergic receptor signaling
    Article Snippet: .. PPAR target gene expression was performed using RT2 Profiler PCR Array Mouse PPAR Targets (Qiagen cat. no. 330231 PAMM-149ZA). .. Western analysis Tissues were homogenized in RIPA buffer (150 mM NaCl, 0.1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 50 mM Tris-HCl pH 8.0) supplemented with protease inhibitor (cOmplete Mini, Roche).

    Article Title: Molecular mechanism of action of bisphenol and bisphenol A mediated by oestrogen receptor alpha in growth and apoptosis of breast cancer cells
    Article Snippet: .. RT-PCR profiler assay kits for apoptosis was used from a commercial vendor which uses 384 well plates to profile the expression of 370 apoptosis related human genes (Qiagen; SABiosciences Corp, Fredrick, MD, USA; Cat#330231 PAHS-3012E). .. Briefly, MCF7 : 5C cells were treated with E2 (10−9 M) for 24, 48 and 72 h or with indicated compounds (in triplicate) for 48 h and total RNA was isolated using the method mentioned earlier.

    Article Title: CHAC2, downregulated in gastric and colorectal cancers, acted as a tumor suppressor inducing apoptosis and autophagy through unfolded protein response
    Article Snippet: .. The human ubiquitination pathway RT2 profiler PCR array The Human Ubiquitination Pathway RT2 Profiler PCR Array (Cat. no. 330231, Qiagen, Hilden, German) was used to profile the expression of 84 key genes involved in the regulated degradation of cellular proteins by the ubiquitin-proteasome. ..

    Article Title: Thyroid hormone induces progression and invasiveness of squamous cell carcinomas by promoting a ZEB-1/E-cadherin switch
    Article Snippet: .. We profiled the expression of 84 genes whose expression is related to the EMT using the human EMT RT² Profiler™ PCR Array (Qiagen). .. Interestingly, most of the EMT genes were modified by D3-depletion (Fig. ).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Qiagen apoptosis related human genes
    Differential effect of bisphenol (BP) and bisphenol A (BPA) on growth and <t>apoptosis</t> of ERα positive breast cancer cells. (A) Dose-dependent effects of BP, BPA and (oestradiol) E2 on growth of MCF7 cells treated for 6 days as indicated. The black bar denotes the level of DNA in vehicle treated cells over a 6-day period. The growth is measured as amount of DNA present in each well. (* P
    Apoptosis Related Human Genes, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 98 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/apoptosis related human genes/product/Qiagen
    Average 99 stars, based on 98 article reviews
    Price from $9.99 to $1999.99
    apoptosis related human genes - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Differential effect of bisphenol (BP) and bisphenol A (BPA) on growth and apoptosis of ERα positive breast cancer cells. (A) Dose-dependent effects of BP, BPA and (oestradiol) E2 on growth of MCF7 cells treated for 6 days as indicated. The black bar denotes the level of DNA in vehicle treated cells over a 6-day period. The growth is measured as amount of DNA present in each well. (* P

    Journal: British Journal of Pharmacology

    Article Title: Molecular mechanism of action of bisphenol and bisphenol A mediated by oestrogen receptor alpha in growth and apoptosis of breast cancer cells

    doi: 10.1111/bph.12122

    Figure Lengend Snippet: Differential effect of bisphenol (BP) and bisphenol A (BPA) on growth and apoptosis of ERα positive breast cancer cells. (A) Dose-dependent effects of BP, BPA and (oestradiol) E2 on growth of MCF7 cells treated for 6 days as indicated. The black bar denotes the level of DNA in vehicle treated cells over a 6-day period. The growth is measured as amount of DNA present in each well. (* P

    Article Snippet: RT-PCR profiler assay kits for apoptosis was used from a commercial vendor which uses 384 well plates to profile the expression of 370 apoptosis related human genes (Qiagen; SABiosciences Corp, Fredrick, MD, USA; Cat#330231 PAHS-3012E).

    Techniques:

    Analysis of CHAC2 expression and its degradation pathway in gastric and colorectal cancer cell lines. ( a ) The expression of CHAC2 in gastric and colorectal cancer cell lines was determined by RT-PCR and western bolt, using GAPDH as a control. ( b ) CHAC2 expression in stably transfected cells confirmed by RT-PCR and western blot. ( c ) Evaluation of CHAC2 expression after treatment of 5-FU or BFA. ( d ) Analysis of the potential degradation pathway of CHAC2 using different inhibitors. AGS-CHAC2 and SW620-CHAC2 cells were treated with 20 μ g/ml CHX with or without MG-132 (20 μ M) or Leu (100 μ M) or CQ (50 mM) in a time course. Cell lysates were subjected to western blot with primary antibodies including CHAC2, p-ERK and GAPDH. ( e ) The effect of ubiquitin on CHAC2 expression. AGS-CHAC2 and SW620-CHAC2 cells were incubated with indicated rhHA-ubiquitin for 12 h, and cell lysates were subjected to western blot with primary antibodies including CHAC2, HA tag andGAPDH. ( f ) 12 maximal upregulated genes checked by the Human Ubiquitination Pathway RT 2 Profiler PCR Array were shown in the bar graph. Among these genes, RNF148 expression significantly increased nearly 1863 times in SW620 as compared to SW48. ( g ) The expression of RNF148 in gastric and colorectal cancer cell lines was determined by RT-PCR. ( h ) Evaluation of RNF148 expression after treatment of 5-FU or BFA. 5-FU, 5-fluorouracil; BFA, Brefeldin A; CHX, Cycloheximide; Leu, leupeptin; CQ, Chloroquine; Ubi, rhHA-ubiquitin

    Journal: Cell Death & Disease

    Article Title: CHAC2, downregulated in gastric and colorectal cancers, acted as a tumor suppressor inducing apoptosis and autophagy through unfolded protein response

    doi: 10.1038/cddis.2017.405

    Figure Lengend Snippet: Analysis of CHAC2 expression and its degradation pathway in gastric and colorectal cancer cell lines. ( a ) The expression of CHAC2 in gastric and colorectal cancer cell lines was determined by RT-PCR and western bolt, using GAPDH as a control. ( b ) CHAC2 expression in stably transfected cells confirmed by RT-PCR and western blot. ( c ) Evaluation of CHAC2 expression after treatment of 5-FU or BFA. ( d ) Analysis of the potential degradation pathway of CHAC2 using different inhibitors. AGS-CHAC2 and SW620-CHAC2 cells were treated with 20 μ g/ml CHX with or without MG-132 (20 μ M) or Leu (100 μ M) or CQ (50 mM) in a time course. Cell lysates were subjected to western blot with primary antibodies including CHAC2, p-ERK and GAPDH. ( e ) The effect of ubiquitin on CHAC2 expression. AGS-CHAC2 and SW620-CHAC2 cells were incubated with indicated rhHA-ubiquitin for 12 h, and cell lysates were subjected to western blot with primary antibodies including CHAC2, HA tag andGAPDH. ( f ) 12 maximal upregulated genes checked by the Human Ubiquitination Pathway RT 2 Profiler PCR Array were shown in the bar graph. Among these genes, RNF148 expression significantly increased nearly 1863 times in SW620 as compared to SW48. ( g ) The expression of RNF148 in gastric and colorectal cancer cell lines was determined by RT-PCR. ( h ) Evaluation of RNF148 expression after treatment of 5-FU or BFA. 5-FU, 5-fluorouracil; BFA, Brefeldin A; CHX, Cycloheximide; Leu, leupeptin; CQ, Chloroquine; Ubi, rhHA-ubiquitin

    Article Snippet: The human ubiquitination pathway RT2 profiler PCR array The Human Ubiquitination Pathway RT2 Profiler PCR Array (Cat. no. 330231, Qiagen, Hilden, German) was used to profile the expression of 84 key genes involved in the regulated degradation of cellular proteins by the ubiquitin-proteasome.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Stable Transfection, Transfection, Incubation, Polymerase Chain Reaction

    Loss of MLK3 induces TNFα-TNFRSF1a axis-mediated apoptosis in CD8 + T cells. (A) Experimental approach for RT 2 PCR array for apoptosis-associated genes. (B) RT 2 PCR array for apoptosis-associated genes in purified CD8 + CD38 + T cells derived from WT and MLK3 −/− mice (pooled, n=3 mice/group). (C) Representative histogram plots (upper panel) and quantification (lower panel) of TNFRSF1a surface expression, gated on CD4 + and CD8 + T cells, respectively. Values are the mean±SD, *p

    Journal: Journal for Immunotherapy of Cancer

    Article Title: Rationalized inhibition of mixed lineage kinase 3 and CD70 enhances life span and antitumor efficacy of CD8+ T cells

    doi: 10.1136/jitc-2019-000494

    Figure Lengend Snippet: Loss of MLK3 induces TNFα-TNFRSF1a axis-mediated apoptosis in CD8 + T cells. (A) Experimental approach for RT 2 PCR array for apoptosis-associated genes. (B) RT 2 PCR array for apoptosis-associated genes in purified CD8 + CD38 + T cells derived from WT and MLK3 −/− mice (pooled, n=3 mice/group). (C) Representative histogram plots (upper panel) and quantification (lower panel) of TNFRSF1a surface expression, gated on CD4 + and CD8 + T cells, respectively. Values are the mean±SD, *p

    Article Snippet: The cDNA was used for RT² Profiler PCR array mouse apoptosis (Qiagen) using real-time PCR (ABI StepOnePlus).

    Techniques: Polymerase Chain Reaction, Purification, Derivative Assay, Mouse Assay, Expressing

    PP2A inhibition decreases radiosensitivity of stem cells in ex vivo organoid cultures. a Shown are optical microscope images of murine intestinal organoids (upper panel) and murine neurospheres (lower panel) untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Pictures have been taken 48 h after IR treatment. b Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl) or PP2A siRNA (siPP2A). Percentage of cells positive for stem cell marker SSEA1 and apoptotic marker CC-3 was calculated from total SSEA1 positive cells. Scoring was performed 4 h after irradiation. c Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Percentage of cells Lgr5-GFP positive was detected by flow cytometry. Scoring was performed 4 h after irradiation. d Murine hematopoietic stem cells were treated with Cal A or left untreated. Cells were irradiated with 2 Gy and apoptosis analyzed after 16 h by Annexin V labeling. e Human neuroprogenitors were treated with Cal A or left untreated. Cells were irradiated with 6 Gy and apoptosis analyzed after 16 h by Annexin V labeling. Scale bar = 100 μm. Error bars indicate SD; * p

    Journal: Cell Death & Disease

    Article Title: Transient PP2A inhibition alleviates normal tissue stem cell susceptibility to cell death during radiotherapy

    doi: 10.1038/s41419-018-0559-0

    Figure Lengend Snippet: PP2A inhibition decreases radiosensitivity of stem cells in ex vivo organoid cultures. a Shown are optical microscope images of murine intestinal organoids (upper panel) and murine neurospheres (lower panel) untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Pictures have been taken 48 h after IR treatment. b Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl) or PP2A siRNA (siPP2A). Percentage of cells positive for stem cell marker SSEA1 and apoptotic marker CC-3 was calculated from total SSEA1 positive cells. Scoring was performed 4 h after irradiation. c Murine intestinal organoid stem cells were left untreated or irradiated at 6 Gy after treatment with Calyculin A (Cal A), control siRNA (siCtrl), or PP2A siRNA (siPP2A). Percentage of cells Lgr5-GFP positive was detected by flow cytometry. Scoring was performed 4 h after irradiation. d Murine hematopoietic stem cells were treated with Cal A or left untreated. Cells were irradiated with 2 Gy and apoptosis analyzed after 16 h by Annexin V labeling. e Human neuroprogenitors were treated with Cal A or left untreated. Cells were irradiated with 6 Gy and apoptosis analyzed after 16 h by Annexin V labeling. Scale bar = 100 μm. Error bars indicate SD; * p

    Article Snippet: To obtain a more holistic view of PP2Ai effects on apoptotic network proteins, we performed RT-PCR analysis on a panel of apoptosis regulatory genes (RT² Profiler™ PCR Array Mouse Apoptosis from Qiagen).

    Techniques: Inhibition, Ex Vivo, Microscopy, Irradiation, Marker, Flow Cytometry, Cytometry, Labeling

    PP2A suppression influences apoptosis signaling. ES cells were treated with Calyculin A (Cal A) or left untreated. Error bars indicate SD; * p

    Journal: Cell Death & Disease

    Article Title: Transient PP2A inhibition alleviates normal tissue stem cell susceptibility to cell death during radiotherapy

    doi: 10.1038/s41419-018-0559-0

    Figure Lengend Snippet: PP2A suppression influences apoptosis signaling. ES cells were treated with Calyculin A (Cal A) or left untreated. Error bars indicate SD; * p

    Article Snippet: To obtain a more holistic view of PP2Ai effects on apoptotic network proteins, we performed RT-PCR analysis on a panel of apoptosis regulatory genes (RT² Profiler™ PCR Array Mouse Apoptosis from Qiagen).

    Techniques: