Figure 6

LPS induces VCAM-1 expression via p300. Cells were pretreated with GR343, and then incubated with LPS for (A) 8 h or (B) 4 h. Levels of VCAM-1 protein (A) and mRNA (B) expression were determined. (C) Cells were pretreated with 1 μM GR343, and then incubated with LPS for 6 h. VCAM-1 promoter activity was determined. (D) Cells were transfected with p300 siRNA, and then incubated with LPS for 8 h. Protein levels of p300 and VCAM-1 were determined. (E) Cells were treated with LPS for the indicated times. The cell lysates were subjected to Western blot using an anti-phospho-p300 antibody. (F) Cells were pretreated with GR343 (GR, 1 μM), PP1 (1 μM), MCI-186 (MCI, 100 μM), APO (100 μM), or SB202190 (SB, 1 μM), and then treated with LPS for 90 min. The cell lysates were subjected to Western blot using an anti-phospho-p300 antibody. (G) Cells were treated with LPS for the indicated times. The cell lysates were subjected to immunoprecipitation using an anti-p300 antibody, and then the immunoprecipitates were analyzed by Western blot using an anti-ATF2 or anti-p300 antibody. All figures are representative of three independent experiments, performed in duplicate. Data are expressed as means ± SEM. *P < 0.05; ^#P < 0.01, as compared with the cells exposed to LPS alone (A, B, C, F), scrambled siRNA alone (D, upper panel), LPS + scrambled siRNA (D, lower panel), or vehicle alone (E, G).