Fig. 3.

The actin cytoskeleton remodelling of gastric cancer (GC) cells is affected by alpha actinin-4 (ACTN4) knockdown. (A) Transmission electron microscopy (TEM) was used to observe the changes of microfilaments in the two GC cell lines. After the actin cytoskeleton remodelling induced by transforming growth factor-β (TGF-β), the number of microfilaments in the short hairpin (sh)ACTN4 group was less than that in the short hairpin negative control (shNC) group, and the arrangement of microfilaments tended to be disordered. Red arrows indicate the cellular microfilaments. (B) Cytoskeleton staining was used to observe the fluorescence and morphological changes of F-actin in the two GC cell lines. After adding TGF-β to induce cell actin skeleton remodelling, F-actin fluorescence was weakened, and the number of pseudopodia was relatively reduced in the shACTN4 group compared with the control group. White arrows indicate cellular filopodia and lamellipodia. (C, D) Western blotting was used to detect the expression levels of ACTN4, phospho-ezrin, phospho-radixin, phospho-moesin, ezrin, radixin and moesin, phospho-vasodilator-stimulated phosphoprotein (VASP) (Ser157), phospho-VASP (Ser239), and VASP in GC cells (C) and the quantified statistical plot drawn after statistical analysis (D). Data are representative of three independent experiments. ^∗P < 0.05, ^∗∗P < 0.01 and ^∗∗∗P < 0.001. ns: no statistical significance; DAPI: 4′,6-diamidino-2-phenylindole; GAPDH: glyceraldehyde-3-phosphate dehydrogenase.