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. 2017 Aug 17;117(8):1164–1175. doi: 10.1038/bjc.2017.260

Figure 1.

Figure 1

FMNL2 targets COMMD10 for ubiquitin degradation in CRC cells. (A) Lysates from SW480 and HCT116 cells were immunoprecipitated with anti-FMNL2 or anti COMMD10 and immunoblotted with the indicated antibodies. (B) Immunofluorescence microscopy analyses of co-localisation of FMNL2 and COMMD10 in SW480 and HCT116 cells. Scale bars represented 50 μm. (C) Lysates from SW480 cells expressing ectopic FMNL2 truncate (525–616a), FMNL2 (617–1092a) or FMNL2 (1–524a) were immunoprecipitated with anti-FLAG and immunoblotted with the indicated antibody. (D) Pull-down of FMNL2 by GST-tagged COMMD10 fragments with indicated boundaries. (E) Pull-down of FH1-wt or FH1-mut of FMNL2- by GST-tagged COMMD10 fragment (1–132a). (F) Schematics outlines of the binding sites of FMNL2 on COMMD10 and COMMD10 on FMNL2 as well. (G) Expressions of FMNL2 and COMMD10 in FMNL2 overexpression or depleting cells by western blotting and were normalised by β-actin expression. (H) Expressions of FMNL2 and COMMD10 in COMMD10 overexpression or depleting cells by western blotting and were normalised byβ-actin expression. (I) Expression of COMMD10 was detected in FMNL2 overexpression SW480 cells before and after MG132 stimulation by western blotting. (J) COMMMD10 was immunoprecipitated from SW480 cell lysates (cells were transfected with FMNL2 or vector control or shFMNL2 or scrambled control) and the recovered material was immunoblotted for ubiquitin (Ubi). (K) Immunofluorescence microscopy analyses of co-localisation of FMNL2 and COMMD10 after MG132 stimulation in HCT116 cells. Scale bars represented 50 μm. A full colour version of this figure is available at the British Journal of Cancer journal online.