Figure 6: MYO7A selectively binds to CASP8.

(a) MYC-MYO7A was co-expressed with V5-CASP8, V5-CASP9 or V5-CASP3 in 293T cells. α-MYC-immunoprecipitation was performed and MYO7A interaction was assessed via immunoblotting. (b) The indicated proteins were co-expressed in 293T cells and the interaction between MYO7A and full-length or deletion mutants of CASP8 was assessed by immunoprecipitation followed by immunoblotting. (c) Confocal microscopy images of SKOV3 cells showing co-localization of MYO7A (green) and CASP8 (red) using the indicated antibodies. Nuclei stained with DAPI (4,6-diamidino-2-phenylindole; blue). (d) Representative confocal microscopy images of proximity ligation assay (PLA) using α-MYO7A and either no α-CASP8 antibody (i) or rabbit α-CASP8 antibody (i–vii) in SKOV3 cells. PLA signals are in green and nuclei are stained with DAPI (blue). PLA between MYO7A and CASP8 on siRNA knockdown of control (non-targeting oligos) (iii) or RIPK1 (iv). Immunoblot analysis of RIPK1 and ACTIN following siRNA knockdown of control or Ripk1. PLA between MYO7A and CASP8 in cells stably expressing CrmA (v), on treatment with 100 nM SM for 5 h (vi), or in response to SM/CrmA treatment (vii).