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. 2018 Nov 2;9(11):1117. doi: 10.1038/s41419-018-1151-3

Fig. 4. TBM induces an AMPK-dependent autophagosome formation.

Fig. 4

a–b Interaction among Beclin1, Atg14L and Bcl-2 in HeLa (a) and SiHa (b) cells was determined by co-immunoprecipitation assay. c–d Interaction between Bcl-2 and Beclin1 in HeLa (c) and SiHa (d) cells was determined by co-immunoprecipitation assay. e HeLa and SiHa cells were transfected with siScramble or siATG5 for 48 h, and then treated with 15 μM TBM for another 24 h. Expression of Atg5 and LC3 was examined by immunoblot. f HeLa and SiHa cells were transfected with siScramble or siBeclin1 for 48 h, and then treated with 15 μM TBM for another 24 h. Expression of Beclin1 and LC3 was examined by immunoblot. g ATP production was detected in cells treated with indicated concentrations of TBM by ATP Assay Kit (Beyotime Biotechnology, S0026). (h) Immunoblot analysis AMPK, ACC, AMPK phosphorylation (Thr172) and ACC phosphorylation (Ser79) in cells treated with the indicated concentrations of TBM for 24 h. β-actin was used as the internal control. i Cells were treated with TBM in the absence or presence of CC. AMPK, AMPK phosphorylation (Thr172) and LC3 were measured by immunoblot. j Cells were treated with TBM in the absence or presence CC for 24 h. LC3 puncta formation was measured by immunofluorescence analysis. Scale bars: 20 μm. k Graph shows quantification of LC3-positive puncta in Figure J. *p < 0.05; **p < 0.01; ***p < 0.001