Participation of PLCγ1 in the feedback monomerization of the EGFR.
(A) ccRICS analysis of PLCγ1 binding to EGFR upon EGF challenge. 2D cross-correlation of eGFP-EGFRwt and mCherry-PLCγ1wt in CHO-K1 cells before (left) and after (right) EGF addition. (B) Cross-correlation index (CCi) between mCherry-PLCγ1, eGFP-EGFR, and their mutants before (white) and after (grey) EGF addition. Lyn-mCherry + eGFP (1) and lyn-mCherry + lyn-eGFP (2) were used as negative controls (CTRL). Data are means ± SD. Symbol * indicates significant difference with p < 0.005 calculated using a two sided T-test with unequal variance. (C) Time traces of diffusion coefficient of eGFP-EGFRY992/1148/1173F, lacking PLCγ binding sites in the upper panel, and eGFP-EGFRwt coexpressed with mCherry-PLCγ1Y783F (inactive mutant, lower panel).