Fig. 5

YAP knockdown facilitated autophagy and autophagy-related cell death in HCC cells. a–d The amount of the autophagy marker LC3B was measured in HCC cell lines (BEL/FU, SK-Hep1, BEL-7402 and HCC-LM3) with YAP knockdown or overexpression under conditions of complete medium or EBSS solution with or without CQ (100 µM) for 6 h. e The autophagosome (yellow puncta) and autolysosome (red puncta) were examined in BEL/FU cells with or without YAP knockdown after transfection of GFP-mRFP-LC3B fusion protein. Left panel, representative images. Right panel, quantification of autophagosomes and autolysosomes within a single cell (from 5 random fields) (scale bar: 25 µm). f–i BEL/FU cells with YAP knockdown were treated with the autophagy inhibitor 3-MA (5 mM) or CQ (100 µM) for 48 h or with the knockdown of ATG5 or BECN1 in the presence of 5-Fu (0.3 mg/ml). All cells were harvested for apoptosis analysis by flow cytometry (F, G) or detection of protein levels of cleaved PARP and cleaved caspase-3 by western blot h, i. β-Actin was used as a loading control. Data are presented as the mean ± SD. **p < 0.01