Fig. 6: Notch3 downregulation induces EGFR dephosphorylation by promoting the endogenous EGFR/PTPH1 interaction. | Oncogenesis

Fig. 6: Notch3 downregulation induces EGFR dephosphorylation by promoting the endogenous EGFR/PTPH1 interaction.

From: NOTCH3 inactivation increases triple negative breast cancer sensitivity to gefitinib by promoting EGFR tyrosine dephosphorylation and its intracellular arrest

Fig. 6

a Raft (R) and non-raft (NR) fractions derived from 6 days of Notch3-silenced cells were used for immunoblot assay with anti-N3EC, anti-pEGFR(Y1173), and anti-EGFR antibodies, to test the effect of Notch3 downmodulation on EGFR-rafts localization. Anti-transferrin and anti-GM1 were used as a fraction markers. b Cells have been activated with EGF ligand for the times indicated, combined or not with Notch3 silencing for 3 days: the expression of phospho-EGFR at tyrosine 1173 and 1068 residues and total EGFR was determined by immunoblot analysis using the specific indicated antibodies. c Control or anti-PTPH1 antibody immunoprecipitates from control and Notch3-silenced cells were probes with anti-EGFR, to detect the EGFR-PTPH1 binding, and with the anti-PTPH1 antibody, to show PTPH1 immunoprecipitated protein levels. The inputs indicated in the panel shows 5% of each total lysate (right panels). Whole cell extracts (WCE) were incubated with anti-N3IC antibody to control the efficiency of Notch3 silencing (left panels). In all panels a, b and c, western blotting against the anti-β-actin was used as a loading control. The results are representative of three independent experiments

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