Fig. 5.

Downregulation of GLYR1 reduces 5-FU-induced apoptosis in CRC cells. a 5-FU was used to induce apoptosis in SW480 and SW620 cells and the expression of p-AKT, p-p38, p-PARP, c-CASP9, c-CASP3 was detected by Western blot analysis. b CCK-8 assays of GLYR1 downregulation on sensitivity of SW480 and SW620 cells to 5-FU; the half maximal inhibitory concentration (IC50) was calculated using GraphPad software. (c-d) Effects of GLYR1 downregulation on apoptosis in SW480 cells induced by 5-FU (8.185 μg/ml) treatment for 48 h was determined by flow cytometric analysis and Hoechst 33258 staining. Representative photographs of Hoechst 33258 staining (× 200, scale = 50 μm); red arrowhead indicates positive apoptotic cells. Error bars represent the mean ± SD of apoptosis (n = 3, n = 5). e Western blot analysis of the expression of the apoptosis-related proteins Bax, PARP, Caspase9, Caspase3 in GLYR1-knockdown SW480 and SW620 cells following 5-FU treatment (8.185 μg/ml, 5.293 μg/ml) for 48 h. Relative expression analysis of cleaved-PARP, cleaved-Caspase9 and cleaved-Caspase3 shown on the right was normalized to the respective pro-protein. Data represent the mean ± SD (n = 3), *P < 0.05, **P < 0.01, ***P < 0.001