Fig. 4

SCAP regulates autophagy in HCC by influencing AMPK signalling. Parental PLC cells (NC) and PLC-SR cells (SR) were treated with sorafenib (6 μm/L) for 24 h. A Ultrastructural analysis of parental cells and PLC-SR cells. The red arrowhead represents autophagic vacuoles (defined to include autophagosomes and autolysosomes) (n = 3). Bar = 50 μm. B Fluorescence and quantification of LC3-positive autolysosomes or autophagosomes in each group (n = 3). Bar = 100 μm. C Immunoblot analysis of LC3 and P62 protein expression in each group (n = 3). Vector cells (Vec) and SCAP knockdown cells (SCAPi) were treated with sorafenib (6 μm/L) for 24 h. D Ultrastructural analysis of SR vector cells and SR SCAP knockdown cells. The red arrowhead represents autophagic vacuoles (defined to include autophagosomes and autolysosomes) (n = 3). E Fluorescence and quantification of LC3-positive autolysosomes or autophagosomes in each group (n = 3). F Immunoblot analysis of SCAP, LC3 and P62 protein expression in each group (n = 3). G Immunoblot analysis of p-AMPK and t-AMPK protein expression in each group (n = 3). SR SCAP knockdown cells were treated with the phosphorylation inhibitor Compound C (SCAPi + CC) (4 μm/L) for 12 h, and Vec cells, SCAPi cells and SCAPi + CC cells were treated with sorafenib (6 μm/L) for 24 h. Immunoblot analysis of p-AMPK, t-AMPK (H), LC3 and P62 (I) protein expression in each group (n = 3). Data are the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. P values were determined by Student’s t test in (A), (B), (C), (D), (E), (F), and (G) and one-way ANOVA in (H) and (I)