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. 2014 Dec 19;22(8):1275–1286. doi: 10.1038/cdd.2014.209

Figure 3.

Figure 3

ERK/MAPK signaling is necessary for anoikis protection. (a) KPL-4 EV and shErbB2 and SUM149 EV and shEGFR cells were plated on poly-HEMA-coated plates for 48 h, and p-ERK levels were measured via immunoblotting. (b and c) KPL-4 and SUM149 cells were plated on poly-HEMA-coated plates in the presence of DMSO or U0126 (10 μM). Caspase activation was measured at 48 h as previously described. Inhibition of the ERK/MAPK pathway was confirmed via western blotting analysis. (d and e) KPL-4 and SUM149 cells were plated in ECM detachment with DMSO or U0126 (10 μM). Cellular viability was measured with the Cell Titer Glo Assay (d) or alamarBlue assay (e). (f and g) KPL-4 (f) and SUM149 (g) cells were plated in attached or detached conditions with DMSO, PD0325901 (1 μM), or LY294002 (25 μM). Caspase activation was measured at 24 h. Cell lysates were prepared and normalized, and protein levels were analyzed via western blotting analysis to confirm inhibitor efficacy. Error bars represent S.E.M. NS, not signifcant