Figure 2. Effects of HDAC inhibitors on CD1d mRNA expression in various tumor cells. (A) A549 cells were treated with TSA (1 μM), SAHA (5 μM), MS-275 (1 μM), NaBu (1 mM), oxaflatin (Oxa, 1 μM), or VPA (1 mM) overnight. Total RNA (2 μg) was used for RT-PCR (upper panel) and total cell lysates were subjected to western blot analysis using antibody specific for acetylated-histone H3 (lower panel). The blot was representative for at least three independent experiments. (B) A549 cells were treated with the indicated dose of TSA or SAHA overnight, or treated with TSA (1 μM) or SAHA (5 μM) for the indicated time. Total RNA (2 μg) was used for RT-PCR. (C) A549, NCI-H292, TC-1 and B16/F0 cells were treated with TSA (1 μM) or SAHA (5 μM) for 16 h, then total RNA (2 μg) were used for RT-PCR. (D) A549 cells were treated with SAHA (3 or 5 μM) or TSA (0.5 or 1 μM) for 16 h. Total RNA were used for real time PCR. Relative CD1d mRNA expression was presented as the mean ± S.D., *:p < 0.05, as compared with basal. Three independent experiments were performed. (E) A549 cells were treated with TSA (1 μM) or SAHA (5 μM) for 24 h, then total cell lysates were subjected to western blot analysis using antibody specific for CD1d and Actin. The blot was representative for at least three independent experiments.