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Figure 3 | Molecular Cancer

Figure 3

From: LIP expression is regulated by IGF-1R signaling and participates in suppression of anoikis

Figure 3

IGF-1 signaling weakly activates EGFR. Western blot analysis of serum starved MCF10A and MCF7 cells stimulated with insulin (10 or 100 nM), IGF-1 (2.6 or 26 nM) or EGF (1.6 or 16 nM) for 10-20 minutes or 16 hours and harvested for analysis of phosphorylation status for the EGFR, IGF-1R, insulin receptor, Akt, or Erk1/2. A). MCF10A whole cell protein extracts, 100 μg for phospho-EGFR (p-EGFR) and 50 μg for total EGFR (EGFR) were analyzed via 7% SDS-PAGE and Western Blotting using anti-EGFR antibodies. B). Upper panel: 1-2 mg of whole cell extracts were immunoprecipitated overnight with anti-EGFR, electrophoresed via 7% SDS-PAGE and immunoblotted with anti-phosphotyrosine (4G10). Lower panel: Whole cell extracts (WCE 50 μg) analyzed via 7% SDS-PAGE and Western Blotting using anti-EGFR antibody. C). Whole cell extracts were immunoprecipitated with 4G10 and immunoblotted with anti IGF-1R or anti-insulin receptor. D). Whole cell protein extracts of MCF10A, 100 μg were analyzed via 7% SDS-PAGE and Western Blotting using specific antibodies to p-EGFR, p-IGF-1R, total EGFR (EGFR), p-Akt, p-Erk1/2, total Akt or total Erk1/2. E). Whole cell protein extracts of MCF7, 100 μg were analyzed via 7% SDS-PAGE and Western Blotting using specific antibodies to p-EGFR, p-IGF-1R, total EGFR (EGFR) and p-Akt, p-Erk1/2, total Akt or total Erk1/2.

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