Fig. 3: Rab27a downregulation inhibits MHCC97H-derived exosome secretion and promotes migration, chemotaxis and invasion of parental cells. | Cell Death & Disease

Fig. 3: Rab27a downregulation inhibits MHCC97H-derived exosome secretion and promotes migration, chemotaxis and invasion of parental cells.

From: HCC-derived exosomes elicit HCC progression and recurrence by epithelial-mesenchymal transition through MAPK/ERK signalling pathway

Fig. 3

a The knockdown efficiency of the Rab27a shRNA was examined by Western blot. b Equal amounts of total exosomal proteins (60 μg) secreted by siRab27a/MHCC97H and siSCR/MHCC97H cells were analysed by Western blot for the presence of the exosomal markers CD63, Alix and TSG101, as well as AFP (a recognised HCC marker). The endoplasmatic reticulum protein GRP78 was used as the negative control. GAPDH was used as loading control. c The total amounts of protein in exosome pellets purified from culture supernatant of MHCC97H (WT), siSCR/MHCC97H and siRab27a/MHCC97H cells were quantified by Bradford assay. d Alteration in migration by Rab27a knockdown was determined by scratch assays. e Alteration in chemotaxis by Rab27a knockdown was measured by chemotaxis assays. f Alteration in invasion by Rab27a knockdown was examined by Matrigel invasion assays. g Alteration in clonogenic ability by Rab27a knockdown was detected by colony formation assays. h Alteration in sphere formation ability by Rab27a knockdown was explored by 3D sphere formation assays. Abbreviation: Exo exosome. *P < 0.05, **P < 0.01 and ***P < 0.001. Scale bar, 1.0 mm. Data are represented as the mean ± S.D. All experiments were repeated at least three times

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