Fig. 5

CHCHD4 regulates the EMT phenotype of tumour cells. a Chart shows GSEA of downregulated proteins detected in SILAC analysis of CHCHD4 (WT)-expressing cells (WT.cl1), compared to control U2OS cells. n = 3. b Volcano plot shows relative expression of all proteins detected in SILAC analysis of CHCHD4 (WT)-expressing cells (WT.cl1) compared to control (Ctrl) U2OS cells [30]. Selected genes highlighted. n = 3. c Western blots show levels of vimentin and N-cadherin in control (Ctrl) U2OS cells, CHCHD4 (WT)-expressing cells (WT.cl1, WT.cl3) and CHCHD4 (C66A/C68A)-expressing cells (C66A/C68A). β-Actin was used as a load control. d Chart shows relative transcript levels of VIM, KRT18 and CHCHD4 detected by Q-PCR in control U2OS cells and CHCHD4 (WT)-expressing cells (WT.cl1). ± SD. n = 3. e Images show a scratch-wound assay of control (Ctrl) U2OS cells, CHCHD4 (WT)-expressing cells (WT.cl1) and CHCHD4 (C66A/C68A)-expressing cells (C66A/C68A) at 0 h and 24 h. White lines denote width of scratch-wound. f Chart shows % wound closure at 24 h relative to 0 h in images shown in (e). ± SD. n = 3. g Immunofluorescence images of vimentin distribution in control U2OS cells and CHCHD4 (WT)-expressing cells (WT.cl1, WT.cl3). h Immunofluorescence images of vimentin distribution in control U2OS cells and CHCHD4 (WT)-expressing cells (WT.cl1) either untreated (NT) or treated with 50 nM rotenone for 72 h