Fig. 2

PHF8 knockdown significantly suppresses proliferation, migration, invasion and autophagy of HCC cells in vitro. a Determination of transfection efficiency of shRNAs targeting PHF8 in SMMC-7721 and Huh7 by qRT-PCR and western-blot assay. Scramble shRNA (shCtrl) was used for negative control. b Inhibited proliferation of SMMC-7721 and Huh7 cells in PHF8 knockdown group by CCK8 assasy (n = 6). c, d) Representative images and quantification of migrated and invasive cells by transwell assay in SMMC-7721 and Huh7 cells (n = 3, magnification, × 100). e Representative immunoblot results of autophagy markers, LC3B and p62 in SMMC-7721 and Huh7 cells with PHF8 knockdown. Both cell lines transfected with indicated shRNAs were cultured in complete medium with 10% FBS or EBSS starvation condition with or without CQ (100 μmol) for 8-h. The ratio of LC3-II to LC3-I and p62 to β-actin were shown at the bottom of each band (n = 3). f Representative fluorescence images of autophagosomes and autolysosomes in SMMC-7721 and Huh7 cells with PHF8 knockdown by tandem mCherry-GFP-LC3 fusion protein assay (magnification, × 400). g Quantification of autophagosomes and autolysosomes from random 5 high-power fields of the merged images of each group. * P < 0.05, ** P < 0.01, *** P < 0.001. Data were presented by mean ± SD