Fig. 7. LLPS of SHP2 mutants recruit and activate SHP2^WT to promote ERK1/2 activation.

(A) Immunoblots of the indicated proteins in HEK293T cells transiently transfected with indicated expression plasmids. (B, C) HEK293T cells were transiently transfected with the indicated amount of SHP2^WT and SHP2^Y297C plasmids. The immunoblots (B) and the densitometry analysis (C) of pERK/ERK levels(means ± SEM, N = 3 experiments) was shown. See Figure S7A for SHP2 knock-out cells. (D) Representative images showing SHP2^WT-mEGFP was readily distributed into SHP2^R498L-mScarlet droplets. Scale bar, 2.5 μm. See also Figure S7C, S7D and S7E.(E) Enzymatic activity (means ± SEM, N = 3 experiments) of indicated proteins under the conditions of LLPS and solution using pNPP as substrate. ***p<0.001. (F) Living images of KYSE520 cells co-expressed with SHP2^WT-mEGFP and SHP2^R498L-mScarlet. Scale bar, 10 μm. See also Figure S7G. (G) Cartoon illustrating that SHP2^mut induces strong LLPS to recruit SHP2^WT and promote ERK activation.