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Fig. 3 | Molecular Cancer

Fig. 3

From: EGFR modulates monounsaturated fatty acid synthesis through phosphorylation of SCD1 in lung cancer

Fig. 3

EGFR mediates phosphorylation of SCD1 at Y14, Y41 and Y55. a 293 T cells were co-transfected with Flag-SCD1 and wildtype or K745R mutant of EGFR-HA for 48 h. Flag-SCD1 was immunoprecipitated and its tyrosine phosphorylation was detected by a pan anti-phosphotyrosine antibody (4G10) in western blot analysis. WT, wild type. b 293 T cells exogenously co-expressing EGFR-HA and Flag-SCD1 were serum-starved overnight, followed by epidermal growth factor (EGF) (50 ng/ml) stimulation for 2 h with/without AG1478 (10 μM) or erlotinib (10 μM). The lysates were subjected to IP/IB with the indicated antibodies. c Lysates from 293 T cells ectopically co-expressing EGFR-HA and wildtype or different mutants of Flag-SCD1 were subjected to IP/IB with the indicated antibodies. d 293 T cells were transfected with wildtype or different mutants of Flag-SCD1 for 48 h. Flag-SCD1 was immunoprecipitated, pre-treated with lambda phosphatase and then incubated with or without a recombinant active EGFR fragment (aa 695–1210). The reaction mixtures were subjected to IB. Top arrow: EGFR fragment auto-phosphorylation; Bottom arrows: tyrosine phosphorylation of Flag-SCD1 and the cleaved product. e HCC827 and H1975 cells were treated with or without erlotinib (1 μM) for 24 h and the lysates were subjected to IP/IB with the indicated antibodies. Densitometry quantification of 4G10/Flag (a-d) and phospho-SCD1 Y55/SCD1 (e) levels is shown

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