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. 2023 Jul 25;14:4467. doi: 10.1038/s41467-023-40212-1

Fig. 6. MET404 independently activates the MET receptor.

Fig. 6

a List of predicted amino acid contacts between MET404 and MET. b Top, illustration of domains on full-length MET. Bottom, 293T cells were transfected with different Flag-tagged MET truncations. Cell lysates were immunoprecipitated with an anti-MET404 antibody and then immunoblotted with an anti-Flag antibody. Representative of three independent experiments. c Protein levels of phospho-MET and downstream phospho-AKT (S473) and phospho-ERK in GSC387 and GSC28 cells after 1 h of stimulation with HGF (100 ng/ml), MET404 (1 µg/ml) or both. Representative of three independent experiments. d Protein levels of phospho-MET and downstream phospho-AKT (S473) and phospho-ERK in GSC456 and GSC23 HGF KO cells after 1 h gradient stimulation of MET404. Representative of three independent experiments. e 293T HGF KO cells were cotransfected with HA- and flag-tagged MET and subjected to coIP analysis. Cell lysates that were stimulated or not stimulated with MET404 (1 µg/ml) were immunoprecipitated with the anti-HA or anti-flag antibody and then immunoblotted with anti-flag or anti-HA antibody. Representative of three independent experiments. f Left, illustration of the NanoBit System. Right, the luminescence of 293T HGF KO cells cotransfected with LgBit- and SmBit-MET was recorded before and after MET404 treatment (1 µg/ml). n = 3 independent experiments. g HGF (100 ng/ml) or MET404 (1 µg/ml) was used to treat METexon2 KO GSC456/GSC23 cells or those overexpressing the flag-tagged MET β subunit. The protein levels of phospho-MET and downstream phospho-AKT (S473) and phospho-ERK were detected in these cells. Representative of three independent experiments. Source data are provided as a Source data file.