Fig. 3

SAHA treatment induces ERK-dependent upregulation of EGR1. a, b and c CME-1 and Yamato-SS cells were exposed to SAHA for the indicated times. In (a) and (c) cells were processed for Western blot analysis with the indicated antibodies. Acetylation of H3 and H4 at K27 and K12, respectively, is shown as marker of HDAC inhibition. In (b) EGR1 mRNA expression was analyzed by qRT-PCR and expressed as relative quantification with respect to untreated cells as calibration sample. Mean relative mRNA values ± SE from three independent experiments are reported. d Effect of 3 h-treatment with SAHA and trametinib, alone and in combination, on ERK activation and EGR1 expression assessed in CME-1 cells by Western blotting. In (a), (c) and (d) actin, tubulin and GAPDH show loading control. Numbers represent the intensity of relevant bands normalized to the respective loading controls