Fig. 3

Cooperative activation of cholesterogenic genes by Ad4BP/SF-1 and SREBP-2 possibly through mutual interaction. a Luciferase reporter gene constructs, Fdps-Luc, Msmo1-Luc, and Dhcr24-Luc, were transfected into Y-1 cells with expression vectors for Ad4BP/SF-1 and SREBP-2. Average values and SDs of the luciferase activities are indicated. Average values in the absence of the expression vectors were normalized to 1.0. n = 4. b Purified FLAG-SREBP-2 protein was mixed with nuclear extracts prepared from Y-1 cells, and then proteins interacting with SREBP-2 were immunoprecipitated with anti-FLAG antibody. The immunoprecipitates were subjected to western blotting using antibodies for Ad4BP/SF-1, FLAG, and CREB. CREB was used as a negative control. Full blot images are shown in Supplementary Fig. 10. NE nuclear extract. c Purified HA-SREBP-2 and FLAG-Ad4BP/SF-1 were mixed, and then immunoprecipitated with anti-FLAG antibody. The immunoprecipitate was subjected to western blotting with anti-HA antibody to examine whether HA-SREBP-2 was co-immunoprecipitated. d A model of cholesterogenic genes regulation is illustrated. Ad4BP/SF-1 possibly regulates the cholesterogenic genes by two ways. Ad4BP/SF-1 directly binds to cholesterogenic genes and then regulates the genes possibly in cooperation with SREBP-2. Besides, Ad4BP/SF-1 indirectly regulates cholesterogenic genes through Srebf2 gene regulation