Fig. 6

T-RECS ASO treatment synergizes with MEKi to inhibit the growth of melanoma cells. a) QRT-PCR analysis shows elevated T-RECS RNA levels in D04 cells after three days of drug-induced Inhibition of MAPK signaling using the MEKi trametinib (20nM or 40nM), when compared to control, treated with DMSO (n = 3). b) D04, MM415 and primary-derived AV5 cells were treated with combinations of T-RECS ASO and trametinib. Combination indices (CI) for dual-treatment effects on cell growth were calculated following the Bliss-model (see methods). Synergistic effects were observed in a range of different concentration combinations in all three cell lines. (n = 2) c) T-RECS ASO treatment significantly inhibited cell growth in the MEKi treatment resistant melanoma cell lines D04RM (p = 0.0002), MM415RM (p = 0.0001), WM3629RM (p = 0.0005) and Sk-Mel-2RM (p = 0.0019). Data were normalized to treatment with non-targeting Control ASO. d) Immunoblotting showing elevated hnRNPA2/B1 protein levels in D04 cells after three days of drug-induced Inhibition of MAPK signaling using the MEKi trametinib (20nM or 40nM), when compared to control, treated with DMSO (n = 3). GAPDH served as a loading control. Significance is shown as p-values calculated by Student’s t-test. * = p < 0.05, ** = p < 0.01, *** = p < 0.001