Skip to main content
. 2021 Oct 30;41(2):191–203. doi: 10.1038/s41388-021-02077-w

Fig. 4. GRB7 conferred resistance to MEKi through regulation of downstream RTK pathway.

Fig. 4

A Western blot analysis showed that combinational inhibition of GRB7 and MEK suppressed the downstream effectors in RTK pathway. HCT116 and SW480 were infected with shRNA4 targeting GRB7 or vector control. After infection, cells were treated with AZD6244 for 24 h. Cell lysates were made for immunoblot analysis with antibodies indicated. ACTIN is as loading control. B Overexpression of GRB7 activated RTK pathway. HCT116 were ectopically expressed GRB7 or vector control. After infection, cells were treated with AZD6244 for 24 h. Cell lysates were made for immunoblot analysis with antibodies indicated. ACTIN is as loading control. C–F Knockdown of GRB7 impaired HGF/EGF-dependent pathway. HCT116 were infected with shRNA4 targeting GRB7 or vector control. After infection, cells were treated with 20 ng/mL HGF(C) or EGF (E) for the indicated time. Cell lysates were made for immunoblot with antibodies indicated. ACTIN is as loading control. D Quantification of (C). F Quantification of (E). G GRB7 interacted with RTKs directly. Cell lysates from GRB7 expressing or vector control cells were subjected to immunoprecipitation with Flag antibody or immunoglobulin G control. The immunoprecipitants (left) or cell lysates (right) were then blotted with the indicated antibodies. H Representative immunofluorescence images of GRB7 in HCT116 treated with 20 ng/mL HGF for indicated time. Scale bar, 20 μM. I Quantification of (H). *p < 0.05, **p < 0.01, ***p < 0.001, or ****p < 0.0001.