Figure 4
Effect of 4bHWE on AKT kinase, SRPK1, SRPK2, and PP1. (a) Huh-7 cells were treated with the indicated concentration of 4bHWE for 24 h. The proteins were analyzed through Western blotting. (b) and (c) Huh-7 cells were treated with 5 μM AKT inhibitor for 1 h and then exposed to 10 μM 4bHWE for 24 h. The proteins and mRNA were analyzed through Western blotting and RT-PCR, respectively. (d) and (e) Huh-7 cells were treated with 20 nM okadaic acid for 1 h and then exposed to 10 μM 4bHWE for 24 h. The proteins and mRNA were analyzed through Western blotting and RT-PCR, respectively. Western blotting: equal amounts of whole cell lysates (20 μg) were separated using SDS-PAGE and immunoblotted with various antibodies as indicated. Tubulin and Histone H3 are shown as internal standards. The fold-change values are presented below each band. RT-PCR: mRNA was extracted and detected for the alternative splicing of the HIPK3, SMAC/DIABLO, and SURVIVIN transcripts. The ratios of the densities of the two bands (alternative exon-containing isoforms to alternative exon-lacking isoforms) are presented below each group. Data are the mean of three independent experiments. *p < 0.05 compared with the control. a p < 0.05 for the comparison between 4bHWE and AKT inhibitor + 4bHWE. b p < 0.05 for the comparison between 4bHWE and okadaic acid + 4bHWE. AI = AKT inhibitor; OA = okadaic acid; M = marker. Full-length blots are presented in Supplementary Fig. S3.
