Fig. 2.

Effects of hindsiipropane B on HIV-1 Tat-mediated expression of HDAC6 and MAPK activation in astrocytes. CRT-MG cells were pretreated with hindsiipropane B for 1 h and then exposed to HIV-1 Tat (50 nM). (A) After 3 h, total RNA was harvested and assayed for mRNA expression of HDAC6 and β-actin by quantitative RT-PCR. (B) After 24 h, cell extracts were collected and evaluated for the levels of HDAC6, acetylated α-tubulin, α-tubulin and β-actin by Western blotting. (C) Cells were pretreated with hindsiipropane B for 1 h and then stimulated with 50 nM HIV-1 Tat for 1 h. Cell lysates were collected and evaluated for the levels of phosphorylated and total ERK, p38, and JNK by Western blotting using relevant antibodies. (D) The intensity of each MAPK bands were quantified by scanning densitometry and normalized to total MAPK levels. Data are shown as mean ± SD of three independent experiments. **P < 0.01, ***P < 0.001, ^#P < 0.05, ^##P < 0.01, ^###P < 0.001, ^^P < 0.01, and ^^^P < 0.001, as compared to the cells treated with HIV-1 Tat alone.