Journal: Journal of Translational Medicine

Article Title: Effect of the mRNA decapping enzyme scavenger (DCPS) inhibitor RG3039 on glioblastoma

doi: 10.1186/s12967-024-05658-x

Figure Lengend Snippet: DCPS inhibition reduced STAT5B expression in GBM cells ( a – b ) GBM cell lines were treated with RG3039 (6 µM) or DMSO for 48 h and then subjected to RNAseq. The top 20 differentially expressed transcription factors were displayed in heatmap ( a ) and volcano plot ( b ). FC, fold change ( c ) Correlation analysis of the expressions of DCPS and STAT5B in GBMs in TCGA database. Statistical analysis by linear regression analysis ( d ) GBM cell lines were transfected with siRNA targeting DCPS for 48 h. The expression of STAT5B mRNA was analyzed by RT-qPCR and normalized to that of β-actin (mean ± SEM, n = 3). Statistical analysis by unpaired t-test, **, p < 0.01, ***, p < 0.001 ( e ) GBM cell lines were treated with RG3039 (6 µM) or DMSO for 48 h. The expression of STAT5B mRNA was analyzed by RT-qPCR and normalized to that of β-actin (mean ± SEM, n = 3). Statistical analysis by unpaired t-test, ****, p < 0.0001 ( f ) GBM cell lines were transfected with siRNA targeting DCPS for 48 h. The expressions of DCPS and STAT5B were analyzed by western blotting ( g ) GBM cell lines were treated with RG3039 (6 µM) or DMSO for 48 h. The expression of STAT5B was analyzed by western blotting

Article Snippet: The membranes were blotted with antibodies against GAPDH (1:5000, 5174 S, CST), DCPS (1:1000, MA5-26131, Invitrogen), or STAT5B (1:1000, 34662 S, CST) at 4 °C overnight.

Techniques: Inhibition, Expressing, Transfection, Quantitative RT-PCR, Western Blot

Journal: Journal of Translational Medicine

Article Title: Effect of the mRNA decapping enzyme scavenger (DCPS) inhibitor RG3039 on glioblastoma

doi: 10.1186/s12967-024-05658-x

Figure Lengend Snippet: The anti-GBM effect of DCPS inhibition was mediated through reducing STAT5B expression ( a ) GBM cell lines with STAT5B overexpression or vector expression were treated with various concentrations of RG3039 or DMSO for 72 h. The IC50 of RG3039 was analyzed by CCK-8 assay ( n = 3) ( b ) Quantification of the ratio of Annexin-V-positive cells in apoptosis assays (see Figure b for details) in GBM cell lines with STAT5B overexpression or vector expression treated with RG3039 (6 µM) to that of cells treated with DMSO (mean ± SEM, n = 3). Statistical analysis by unpaired t-test, **, p < 0.01, ***, p < 0.001 ( c ) Representative images of colony formation assays in GBM cell lines with STAT5B overexpression or vector expression treated with various concentrations of RG3039 or DMSO for 14 days ( d ) Quantification of the ratio of colony formation of GBM cells with STAT5B overexpression or vector treated with RG3039 to cells treated with DMSO (mean ± SEM, n = 3). Statistical analysis by unpaired t-test, NS, no significance, *, p < 0.05, ***, p < 0.001 ( e ) Schematic diagram summarizing that RG3039 inhibits DCPS regulating the expression of STAT5B, which plays an essential role in tumor progression

Article Snippet: The membranes were blotted with antibodies against GAPDH (1:5000, 5174 S, CST), DCPS (1:1000, MA5-26131, Invitrogen), or STAT5B (1:1000, 34662 S, CST) at 4 °C overnight.

Techniques: Inhibition, Expressing, Over Expression, Plasmid Preparation, CCK-8 Assay