Figure 4

Endogenous CBP represses CLOCK/BMAL1-mediated transcription in NIH3T3 cells. (A) Upper panel: Cells grown in six-well plates were transiently transfected with either pSUPER or pSUPER-CBP (1.8 μg), together with either pCMV-HA or pHA-CBP1-1098 (200 ng), as indicated. Exogenous CBP was detected by Western blotting using anti-HA antibody. Lower panel: Cells grown in six-well plates were transiently transfected with either pSUPER or pSUPER-CBP (1.8 μg). Endogenous CBP was detected by Western blotting using anti-CBP antibody. α-tubulin serves as an equal loading control. (B) Cells were transiently transfected with either pE-less or pE-box (2 ng), together with either pSUPER or pSUPER-CBP (100 ng), and with or without pcDNA3CLOCK (100 ng) plus pcDNA3BMAL1 (100 ng) alone or in combination with pcDNA3CBP (50 ng), as indicated. Empty vector (pcDNA3) was used to standardize the total amount of transfected DNA (502 ng). An asterisk indicates p < 0.05 (Student's t test, n = 9).