Extended Data Fig. 5: Recombinant C-E-cad activates EGFR. (Related to Fig. 5).
a, IB. rC-E-cad with different concentrations were used to treat indicated cells. EGF was used as a positive control. b, IB for GSCH2S and GSC17 with indicated modifications. c, The percentage of SA-β-Gal positive cells of GSCH2S and GSC17 E-cadherin KO with indicated modification. d, IB of GSCH2S and GSC17 E-cadherin KO with indicated modification. e, Mass spectrometry analysis identified EGFR peptide sequences that was pulled down by the anti-C-E-cad antibody. (IB for GSC387 and 4121 with indicated modifications were separately treated with or without rC-E-cad proteins.). g, ELISA of IL-6 level in GSC387 and 4121 with indicated modifications. h, IB. Left, GSC4121 with stable KD of EGFR, Met, PDGFRA, or IL6R and treated with or without rC-E-cad (200 ng/ml). Right. GSC4121 with stable EGFR KD were re-expressed with WT EGFR or EGFR with a Y1068A mutation, then treated with rC-E-cad. i, IB. non-GSC387 and 4121 were modified or treated as indicated. j, LDA assay for non-GSC387 and 4121 with indicated modification or treatments. k, IB. EGFR-HIS, GST-C-E-Cad and GST-C-E-Cad-△14aa was purified and GST pull down assay was applied. l, Docking analysis of C-E-cad and EGFR CR2 domain. m, Tumor volume in GSC387, 4121 and H2S with indicated modifications. n, Two-sided, Log-rank analysis of mice intracranially implanted with GSC387, 4121 and H2S with indicated modifications, ***p < 0.001o. Tumor volumes and Two-sided, Log-rank analysis of mice bearing indicated GSC tumor xenografts with 5 mice per group. ***p < 0.001In a-k, n = 3 independent experiments. In m-o, n = 5 independent experiments. In c,g,j,m.o, data were shown as mean ± SD, two-sided t test,***p < 0.001.
