Fig. 2

Fisetin and RSK inhibitors have similar effect on YB-1 phosphorylation. A The indicated cells were treated with fisetin (75 µM), LJI308 (LJI) (2.5 µM) or BI-D1870 (BID) (2.5 µM). Cytoplasmic and nuclear protein fractions were isolated after 24 h and subjected to SDS-PAGE. Phospho-YB-1, total YB-1, phospho-AKT and total AKT1 were detected by Western blotting. α-Tubulin and lamin A/C were used as cytoplasmic and nuclear markers, respectively. Actin was detected as loading control. B The histograms represent the mean densitometry values ± SD of P-YB-1 to actin and YB-1 to actin from 3 independent experiments normalized to DMSO treated control (Ctrl) condition. The asterisks indicate significant fisetin mediated changes on YB-1 phosphorylation (*p < 0.05, **p < 0.01, ***p < 0.001), ****p < 0.0001; students t-test). C Cells were treated with and without fisetin (75 µM, 24 h) and mock irradiated or irradiated 4 Gy. Cytoplasmic and nuclear protein fractions were isolated at the indicated times after IR and subjected to SDS-PAGE. Phospho-YB-1 and total YB-1 were detected by Western blotting. α-Tubulin and lamin A/C were used as cytoplasmic and nuclear markers, respectively. Actin was detected as loading control