Identification of Gsdf-Ncoa5 interaction via yeast two-hybrid assay and co-immunoprecipitation.

(A) Confirmation of 34 candidate interacting proteins of Gsdf-A via yeast two-hybrid assay. +: pGBKT7–53 and pGADT7-T co-transformed as positive control; −: pGBKT7-Lam and pGADT7-T co-transformed as negative control. (B) List of the 27 potential interaction partners of Gsdf-A. (C-D) qPCR analysis of cyp19a1a expression in the CAB cells transfected with different plasmids. Different letters represent statistical differences (P<0.01). The highest expression level in each qPCR analysis was used as control and defined as 1. (E) Co-IP of Gsdf-A-Flag and Gsdf-B-Flag with Ncoa5-HA in CAB cells transfected with the indicated plasmids. Anti-Flag Ab was used for Co-IP. Sf1: Sf1-A and Sf1-B; Foxl2b: Foxl2b-A and Foxl2b-B; Ncoa5: Ncoa5-B.