mdl29951 treatment  (MedChemExpress)

Journal: Cell Death & Disease

Article Title: G-protein-coupled receptor GPR17 inhibits glioma development by increasing polycomb repressive complex 1-mediated ROS production

doi: 10.1038/s41419-021-03897-0

Figure Lengend Snippet: A Control and U87-GPR17 cells were subjected to RNA-Seq analysis. Volcano plot depicted gene expression changes between control and U87-GPR17 cells. B Control and U87-shGPR17 cells were subjected to RNF2 ChIP-Seq analysis. Density heatmap showed the RNF2 recruitment within ±2 kb around the RNF2 peak center. C Venn diagram showing the overlapped genes in ChIP-Seq and RNA-Seq analyses. D Gene list ranked by RNA-Seq p value with RNF2 binding on promoter-TSS region. E Diagram showing RNF2 enrichment on KLF9 promoter-TSS region. F – H U87-shScr/shGPR17 ( f ), U87-Vec/GPR17 ( G ), or U87MG cells treated with Vehicle/MDL29951 (300 μM) for 48 h ( H ) were subjected to RNF2 and H2AK119ub ChIP assay; real-time PCR analysis was then performed to assess the relative enrichment of RNF2 and H2AK119ub in promoter-TSS region of KLF9 . I Correlation plot of RNF2 and KLF9 mRNA expression in glioma, grades II–IV, from the TCGA dataset ( n = 667). J , K Control, U87/U251-GPR17, or U87/U251-shGPR17 cells were prepared as in Fig. . Cells were harvested for real-time PCR analysis to assess the mRNA levels of RNF2 and KLF9 . L Control or U87/U251-GPR17 cells were transfected with control or RNF2-overexpressing vectors for 48 h, and then subjected to real-time PCR analysis to assess the mRNA levels of KLF9 . For all panels, data represent the means ± SEM from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, Student’s t- test.

Article Snippet: For NAC treatment, U87MG or U251 cells were treated with vehicle or 5 mM N -acetyl- l -cysteine (Sigma, Cat# A8199) for 48 h. For PRT4165 treatment, cells were treated with vehicle or 10 μM PRT4165 (MCE, Cat# HY-19817) for 48 h. For MDL29951 treatment, cells were treated with vehicle or 300 μM MDL29951 (MCE, Cat# HY-16312) for 48 h. Ten microliters of Cell Counting Kit-8 (MCE, Cat# HY-K0301) was then added to each well and the cells were incubated for 2 h at 37 °C.

Techniques: Control, RNA Sequencing Assay, Expressing, ChIP-sequencing, Binding Assay, Real-time Polymerase Chain Reaction, Transfection

Journal: Cell Death & Disease

Article Title: G-protein-coupled receptor GPR17 inhibits glioma development by increasing polycomb repressive complex 1-mediated ROS production

doi: 10.1038/s41419-021-03897-0

Figure Lengend Snippet: A U87MG and U251 cells treated with vehicle or MDL29951 (300 μM) for 48 h, and CCK-8 assay was performed to examine viable cell numbers. Data represent the means ± SEM from three independent experiments. B – D BALB/c nude mice with subcutaneously xenotransplanted tumor were intraperitoneally injected with 10 mg/kg MDL29951 for 16 days, and then tumor samples were collected for the measurements of tumor sizes ( B ), volumes ( C ), and weights ( D ), ( n = 7). E H&E staining of sections from subcutaneously xenotransplanted tumors. Scale bar 10 μm. F , G Immunofluorescent staining against Ki67 ( F ) and TUNEL ( G ) of the tumor sections, scale bar 50 μm. H IHC staining against p-PKA or RNF2 of the subcutaneously xenotransplanted tumor. Scale bar 50 μm. I A diagram depicting the working model of the inhibitory effect of GPR17 on glioma tumorigenesis. For all panels, * p < 0.05, *** p < 0.001, Student’s t- test.

Article Snippet: For NAC treatment, U87MG or U251 cells were treated with vehicle or 5 mM N -acetyl- l -cysteine (Sigma, Cat# A8199) for 48 h. For PRT4165 treatment, cells were treated with vehicle or 10 μM PRT4165 (MCE, Cat# HY-19817) for 48 h. For MDL29951 treatment, cells were treated with vehicle or 300 μM MDL29951 (MCE, Cat# HY-16312) for 48 h. Ten microliters of Cell Counting Kit-8 (MCE, Cat# HY-K0301) was then added to each well and the cells were incubated for 2 h at 37 °C.

Techniques: CCK-8 Assay, Injection, Staining, TUNEL Assay, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: G-protein-coupled receptor GPR17 inhibits glioma development by increasing polycomb repressive complex 1-mediated ROS production

doi: 10.1038/s41419-021-03897-0

Figure Lengend Snippet: A Control and U87-GPR17 cells were subjected to RNA-Seq analysis. Volcano plot depicted gene expression changes between control and U87-GPR17 cells. B Control and U87-shGPR17 cells were subjected to RNF2 ChIP-Seq analysis. Density heatmap showed the RNF2 recruitment within ±2 kb around the RNF2 peak center. C Venn diagram showing the overlapped genes in ChIP-Seq and RNA-Seq analyses. D Gene list ranked by RNA-Seq p value with RNF2 binding on promoter-TSS region. E Diagram showing RNF2 enrichment on KLF9 promoter-TSS region. F – H U87-shScr/shGPR17 ( f ), U87-Vec/GPR17 ( G ), or U87MG cells treated with Vehicle/MDL29951 (300 μM) for 48 h ( H ) were subjected to RNF2 and H2AK119ub ChIP assay; real-time PCR analysis was then performed to assess the relative enrichment of RNF2 and H2AK119ub in promoter-TSS region of KLF9 . I Correlation plot of RNF2 and KLF9 mRNA expression in glioma, grades II–IV, from the TCGA dataset ( n = 667). J , K Control, U87/U251-GPR17, or U87/U251-shGPR17 cells were prepared as in Fig. . Cells were harvested for real-time PCR analysis to assess the mRNA levels of RNF2 and KLF9 . L Control or U87/U251-GPR17 cells were transfected with control or RNF2-overexpressing vectors for 48 h, and then subjected to real-time PCR analysis to assess the mRNA levels of KLF9 . For all panels, data represent the means ± SEM from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, Student’s t- test.

Article Snippet: For NAC treatment, U87MG or U251 cells were treated with vehicle or 5 mM N -acetyl- l -cysteine (Sigma, Cat# A8199) for 48 h. For PRT4165 treatment, cells were treated with vehicle or 10 μM PRT4165 (MCE, Cat# HY-19817) for 48 h. For MDL29951 treatment, cells were treated with vehicle or 300 μM MDL29951 (MCE, Cat# HY-16312) for 48 h. Ten microliters of Cell Counting Kit-8 (MCE, Cat# HY-K0301) was then added to each well and the cells were incubated for 2 h at 37 °C.

Techniques: Control, RNA Sequencing Assay, Expressing, ChIP-sequencing, Binding Assay, Real-time Polymerase Chain Reaction, Transfection

Journal: Cell Death & Disease

Article Title: G-protein-coupled receptor GPR17 inhibits glioma development by increasing polycomb repressive complex 1-mediated ROS production

doi: 10.1038/s41419-021-03897-0

Figure Lengend Snippet: A U87MG and U251 cells treated with vehicle or MDL29951 (300 μM) for 48 h, and CCK-8 assay was performed to examine viable cell numbers. Data represent the means ± SEM from three independent experiments. B – D BALB/c nude mice with subcutaneously xenotransplanted tumor were intraperitoneally injected with 10 mg/kg MDL29951 for 16 days, and then tumor samples were collected for the measurements of tumor sizes ( B ), volumes ( C ), and weights ( D ), ( n = 7). E H&E staining of sections from subcutaneously xenotransplanted tumors. Scale bar 10 μm. F , G Immunofluorescent staining against Ki67 ( F ) and TUNEL ( G ) of the tumor sections, scale bar 50 μm. H IHC staining against p-PKA or RNF2 of the subcutaneously xenotransplanted tumor. Scale bar 50 μm. I A diagram depicting the working model of the inhibitory effect of GPR17 on glioma tumorigenesis. For all panels, * p < 0.05, *** p < 0.001, Student’s t- test.

Article Snippet: For NAC treatment, U87MG or U251 cells were treated with vehicle or 5 mM N -acetyl- l -cysteine (Sigma, Cat# A8199) for 48 h. For PRT4165 treatment, cells were treated with vehicle or 10 μM PRT4165 (MCE, Cat# HY-19817) for 48 h. For MDL29951 treatment, cells were treated with vehicle or 300 μM MDL29951 (MCE, Cat# HY-16312) for 48 h. Ten microliters of Cell Counting Kit-8 (MCE, Cat# HY-K0301) was then added to each well and the cells were incubated for 2 h at 37 °C.

Techniques: CCK-8 Assay, Injection, Staining, TUNEL Assay, Immunohistochemistry