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. 2018 Feb 28;7(2):23. doi: 10.1038/s41389-017-0015-5

Fig. 6. Combined Src, MEK1/2 and AKT/P70S6K inhibition more effectively reduces rpS6 phosphorylation and potentiates inhibitor effects.

Fig. 6

a Western blot analysis was performed on whole cell lysates from the 8505C and C643 cell lines treated with either DMSO, 100 nM trametinib, 100 nM dasatinib, dasatinib+trametinib, 2.5 μM AT7867, trametinib+AT7867, dasatinib +AT7867, or trametinib+dasatinib+AT7867 for 4 h and analyzed for the indicated antibodies. b Clonogenic growth was detected by crystal violet staining in the 8505C or C643 cell lines. The cells were treated with either DMSO, 100 nM trametinib, 100 nM dasatinib, 2.5 μM AT7867, trametinib+AT7867, dasatinib+AT7867, or trametinib+dasatinib+AT7867 for 3 days. Following 3 days of treatment, the cells were released for an additional 6 days. Colony area signal intensity was measured using Odyssey CLx imager (Li-Cor), and presented as percent fold change relative to the DMSO-treated wells. Data as means±SEM (n = 2). c Cleaved caspase 3/7 activity was measured after an 8 h incubation with either DMSO, 100 nM trametinib, 100 nM dasatinib, dasatinib+trametinib, 2.5 μM AT7867, trametinib+AT7867, dasatinib+AT7867, or trametinib+dasatinib+AT7867 in the 8505C or C643 cell line. Data as means±SEM (n = 3; Student’s t-test; *P < 0.05)