(A) Physical interaction between CAR and Baf60a in transiently transfected 293T cells. IP, immunoprecipitation; Myc-Baf60a, Myc-tagged Baf60a; HA-CAR, HA-tagged CAR.
(B) Immunoblots of immunoprecipitated proteins or total protein lysates from HepG2 cells transduced with adenoviruses expressing Flag/HA-tagged CAR (FH-CAR). Note that FH-CAR interacts with endogenous SWI/SNF subunits.
(C-D) Relative enrichment of Baf60a (C) and CAR (D) at the putative CAR binding site on the proximal CAR promoter. RE1, CAR response element 1; RE2, CAR response element 2.
(E) ChIP assay with chromatin lysates prepared from flox/flox and BaLKO mouselivers using indicated antibodies.
(F) ChIP assay with chromatin lysates prepared from primary hepatocytestransduced with GFP or Baf60a adenoviruses.
(G-H) WD-fed flox/flox and BaLKO mice were transduced with adenoviruses expressing GFP or CAR for 1 week. Shown are qPCR analysis of hepatic gene expression (G) and measurement of plasma cholesterol levels (H); *P < 0.05, BaLKO GFP vs. flox/flox GFP; #P < 0.05, BaLKO CAR vs. BaLKO GFP. Data in C-F represent mean ± s.d. and are representative of three independent experiments; Data in G and H represent mean ± s.e.m. (n = 4-5); #P < 0.05, and *P < 0.05 by two-tailed Student's t-test.
See also Figure S3.