Figure 7
From: CBP/p300 is a cell type-specific modulator of CLOCK/BMAL1-mediated transcription
CLOCK and BMAL1 do not coimmunoprecipitate with CBP. (A) NIH3T3 cells were transiently transfected with pmyc-YFP-CREBDIEDML (1 μg), in the presence or absence of pcDNA3CBP (2 μg). Empty vector (pcDNA3) was used to standardize the total amount of transfected DNA (3 μg). (B) NIH3T3 cells were transiently transfected with pcDNA3CBP (2 μg), together with either pCMV-myc (6 μg), pmyc-CLOCK (6 μg), pmyc-BMAL1 (6 μg) or pmyc-YFP-CREBDIEDML (1 μg) plus pCMV-myc (5 μg). (C) NIH3T3 cells were transiently transfected with either pCMV-myc (6 μg), pmyc-CLOCK (6 μg), pmyc-BMAL1 (6 μg) or pmyc-YFP-CREBDIEDML (1 μg) plus pCMV-myc (5 μg). (D) COS-1 cells were transiently transfected with pcDNA3CBP (1 μg), together with either pCMV-myc (3 μg), pmyc-CLOCK (3 μg), pmyc-BMAL1 (3 μg) or pmyc-YFP-CREBDIEDML (0.5 μg) plus pCMV-myc (2.5 μg). (E) COS-1 cells were transiently transfected with either pCMV-myc (3 μg), pmyc-CLOCK (3 μg), pmyc-BMAL1 (3 μg) or pmyc-YFP-CREBDIEDML (0.5 μg) plus pCMV-myc (2.5 μg). Immunoprecipitations (IP) was performed using anti-rabbit IgG or anti-CBP antibodies, and then Western blot analysis was performed using anti-myc or anti-CBP antibodies, as indicated. The input lanes represent 10% of total cell lysates in the binding reaction.