Fig. 1: CENTLEIN interacts with both SUN5 and PMFBP1.

a A candidate-based approach by identification of SUN5-binding centrosomal proteins. FLAG-SUN5 and one of the GFP-tagged plasmid, including the empty vector, GFP-CENTLEIN, GFP-CEP68, GFP-BBS4, GFP-CP110, GFP-NEK2A, GFP-CPAP, GFP-CETN2, and GFP-PLK1, were co-transfected into HEK293T cells. Twenty-four hours after transfection, cells were collected for immunoprecipitation (IP) with anti-GFP antibody and analyzed with FLAG and GFP antibodies, respectively. Red asterisks indicate GFP-tagged proteins. b, c CENTLEIN could bind with SUN5. Empty vector, GFP-CENTLEIN and FLAG-SUN5 were co-transfected into HEK293T cells. Twenty-four hours after transfection, cells were collected for immunoprecipitation (IP) with anti-GFP antibody (b) or anti-FLAG antibody (c) and analyzed with FLAG and GFP antibodies, respectively. Asterisk indicates IgG heavy chains. d, e CENTLEIN could interact with PMFBP1. Empty vector, MYC-CENTLEIN, and GFP-PMFBP1 were co-transfected into HEK293T cells. Twenty-four hours after transfection, cells were collected for immunoprecipitation (IP) with anti-MYC antibody (d) or anti-GFP antibody (e) and analyzed with MYC and GFP antibodies, respectively. f, g CENTLEIN could not interact with SPATA6. Empty vector, GFP-CENTLEIN, and FLAG-SPATA6 were co-transfected into HEK293T cells. Twenty-four hours after transfection, cells were collected for immunoprecipitation (IP) with anti-FLAG antibody (f) or anti-GFP antibody (g) and analyzed with FLAG and GFP antibodies, respectively. Asterisk indicates IgG heavy chains. The experiment was repeated three times independently with similar results (a–g).