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. 2013 Jul 31;8(7):e68966. doi: 10.1371/journal.pone.0068966

Figure 1. Mig6 is upregulated in an erlotinib resistant cell line which suppresses EGFR phosphorylation.

Figure 1

A) Erlotinib-sensitive (SCC-S) and -resistant (SCC-R) cells were treated with erlotinib and cell viability was assayed. Values were set at 100% for untreated controls. B) Immunoblot analysis of protein expression in SCC-S and -SCC-R cell lines. C) SCC-S and SCC-R cells were treated with EGF at the indicated times and Mig6 protein expression was analyzed. D) Mig6 mRNA expression was examined by real-time quantitative PCR after EGF treatment at the indicated times. Mig6 mRNA expression was normalized to GAPDH expression. E) SCC-S and SCC-R cells were serum-stripped and stimulated with EGF for 60 min. Immunoprecipitation (IP) was performed against EGFR, followed by immunoblotting against Mig6 and EGFR. F) Densitometric quantification of Mig6 and EGFR. Data are presented as the ratio of Mig6/EGFR to indicate how many Mig6 molecules are associated with each EGFR molecule. All ratios are presented in relative arbitrary values. G) SCC-R cells were transfected with either scrambled siRNA or siRNA targeting Mig6 for 48 hrs. Cells were stripped in serum free medium overnight and stimulated with EGF for 15 or 60 min.