Figure 8: Interrupting the TRB3/p62 interaction inhibits tumour proliferation and invasion.

(a) Wound-healing assay of HepG2 cells treated with Pep2–A2 or Pep2-con plus or minus IGF-1 at indicated times. (b,c) Pep2–A2 attenuates IGF-1-induced invasion of HepG2 cells. Data are representative transwell; scale bar, 100 μm (b). 3D Matrigel culture assay, scale bar, 100 μm (c). (d) Pep2–A2 attenuates IGF-1-induced proliferation of HepG2 cells. Data are representative images of Edu labelling. Scale bar, 18.75 μm. (e,f) A p62^mut expression vector that lacks the C-terminal LIR and UBA domains was constructed. HepG2 cells were trasfected with p62-siRNA alone or together with p62^mut expression vector. After 48 h of trasfection, cells were treated with Pep2–A2 or Pep2-con (5 μM) for 24 h, and then cell proliferation and invasion activities were evaluated through Edu (e) and transwell (f) assays. Scale bar, 18.75 μm (e) and 500 μm (f). (g,h) HepG2 cells were treated with Pep2-con, Pep2–A2 or Pep2–A2^mut (5 μM) for 24 h. The proliferation and invasion activities were evaluated with Edu (g) or transwell (h) assay. For all panels, n=3 independent experiments. Scale bar, 18.75 μm (g) and 500 μm (h). Data indicate mean±s.e.m. Statistical significance was determined with one-way ANOVA; **P<0.01; ***P<0.001.