Figure 1

Isoproterenol signaling increases HDAC6 expression in lung cancer cells. (a) Effects of isoproterenol on the expression levels of type II HDACs in H1299 human lung cancer cells. (b) Effects of cAMP signaling on the expression of HDAC6. (c) Temporal patterns of the expression levels of HDAC6 protein and mRNA following isoproterenol treatment. (d) Effects of transcription inhibition on the isoproterenol-induced increase in HDAC6 expression. H1299 cells were pretreated with actinomycin D (8 μM) for 2 h before isoproterenol treatment (20 μM), and the cells were harvested at 30 h for analyses of the mRNA levels and at 48 h for the western blotting analysis. H1299 human lung cancer cells were treated with 20 μM isoproterenol (ISO), 20 μM forskolin or 20 μM PGE₂ and harvested at 48 h or at the indicated time points for western blotting analysis. β-Actin was analyzed as a loading control. The western blotting band densities were quantified using the Multi Gauge v.2.3 software and are expressed as the ratio relative to the control band density. The HDAC6 mRNA level was analyzed by quantitative real-time PCR and normalized to GAPDH mRNA and is expressed as the ratio relative to the control level. The filled bar represents the HDAC6 protein, and the empty bar represents the HDAC6 mRNA. Asterisks (*) indicate statistically significant differences compared with the respective control (P<0.05, Mann–Whitney U-test).