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. 2013 Aug 26;110(37):14924–14929. doi: 10.1073/pnas.1303640110

Fig. 5.

Fig. 5.

Expression of monobodies decouples GAB2 and SHP2 and modulates downstream signaling. (A) The interaction of endogenous SHP2 and GAB2 was monitored by reciprocal immunoprecipitation experiments on overexpression of monobodies and BCR-ABL. The bar graph quantifies the SHP2–GAB2 interaction and its perturbation by monobody expression from six (for GAB2) and two (for SHP2) independent immunoprecipitation experiments. ns, not significant. *P < 0.05; **P < 0.005. (B) The interaction of endogenous SHP2 with GAB2 in the presence of monobodies was studied in HEK293 cells stably overexpressing HA-tagged GAB2. (C and D) HEK293 cells were transiently transfected with the indicated expression constructs, and total cell lysates were immunoblotted with the indicated antibodies. The bar graphs quantify the phosphorylation of paxillin (C) and STAT5 (D) and its perturbation by monobody expression (n = 2). (E) HCC1171 cells stably expressing the TAP-tagged monobody clones and total cell lysates immunoblotted with the indicated antibodies.