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. Author manuscript; available in PMC: 2013 Mar 5.

Published in final edited form as: Gastroenterology. 2010 Apr 29;139(2):598–608. doi: 10.1053/j.gastro.2010.04.046

(A) Treatment of S2-013 cells with a pool of either atg5 siRNA or beclin1 siRNA shows a significant decrease in Atg5 or Beclin1 levels after 24 h and 48 h, which is absent in cells treated with lipofectamine alone (control) or with non-silencing siRNA (NS). Actin is used as a loading control. (B, C) Treatment of S2-013 with 200 nM triptolide for 24 h following a knock-down of atg5 or beclin1 genes with siRNA pool for 48 h shows a significant decrease in LC3 punctate pattern as compared to cells treated with non-silencing siRNA or lipofectamine alone followed by triptolide. The white dotted line indicates the outline of the cells. S2-013 and S2-VP10 cells treated with triptolide 200 nM for 48 h following atg5 or beclin1 knock-down shows a decrease in cell viability which is comparable to control or NS cells treated with triptolide (D). Knock-down of atg5 or beclin1 followed by triptolide 200 nM treatment for 48 h (E) or 24 h (F) shows a significant increase in caspase-3 activation and Annexin V positive cells when compared to control or NS cells treated with triptolide. The data shown are representative of three independent experiments. The bars represent mean ± SEM, n=3, * p<0.05, **p <0.01 (t test).

(A) Treatment of S2-013 cells with a pool of either atg5 siRNA or beclin1 siRNA shows a significant decrease in Atg5 or Beclin1 levels after 24 h and 48 h, which is absent in cells treated with lipofectamine alone (control) or with non-silencing siRNA (NS). Actin is used as a loading control. (B, C) Treatment of S2-013 with 200 nM triptolide for 24 h following a knock-down of atg5 or beclin1 genes with siRNA pool for 48 h shows a significant decrease in LC3 punctate pattern as compared to cells treated with non-silencing siRNA or lipofectamine alone followed by triptolide. The white dotted line indicates the outline of the cells. S2-013 and S2-VP10 cells treated with triptolide 200 nM for 48 h following atg5 or beclin1 knock-down shows a decrease in cell viability which is comparable to control or NS cells treated with triptolide (D). Knock-down of atg5 or beclin1 followed by triptolide 200 nM treatment for 48 h (E) or 24 h (F) shows a significant increase in caspase-3 activation and Annexin V positive cells when compared to control or NS cells treated with triptolide. The data shown are representative of three independent experiments. The bars represent mean ± SEM, n=3, * p<0.05, **p <0.01 (t test).