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. 2014 Dec 11;5(12):e1571. doi: 10.1038/cddis.2014.540

Figure 6.

Figure 6

Inhibition of siBMPR2-induced autophagy sensitized chondrosarcoma cells to siBMPR2-induced apoptotic cell death. (a) Pretreatment of cells with 3-MA or siBeclin-1 sharply reduced the number of viable siBMPR2-treated cells, as assayed by MTS. (b) Colony formation of chondrosarcoma cells was decreased more sharply by Beclin-1 and BMPR2 siRNA transfection than by BMPR2 siRNA. (c) SW1353 cells were stained with Annexin-V-FITC (20 mg/ml) and PI (20 mg/l) following siBMPR2 treatment with or without siBeclin-1 transfection for 48 h. Apoptosis was analyzed by flow cytometry. (d) Compared with siBMPR2 transfection group, the bar graph showed a significant increase in apoptosis rate in co-transfection of siBMPR2 and siBeclin-1. (e) Apoptosis- and autophagy-related proteins were examined by western blot following siBMPR2 transfection with or without siBeclin-1 for 48 h. (f) Compared with siBMPR2 transfection group, the bar graph showed a significant increase in cleaved PARP in co-transfection of siBMPR2 and siBeclin-1 (P<0.01). (g) The bar graph showed a great decrease in LC3-II in co-transfection of siBMPR2 and siBeclin-1 compared with siBMPR2 transfection group (P<0.01). Representative data from one of three independent experiments are shown in (b–e)