Fig. 2: ESR1 mutations induced a global kinome reprogramming.

a Flowchart demonstrating the total number of kinases captured in KiP experiment and the selection of kinases used to perform KEGG pathway analysis. Hyperactivated was defined as Y537S/WT ≥ 1.5-fold. Unique was defined as having corresponding peptide detection by MS in only MCF-7 ESR1 WT or MCF-7 ESR1 Y537S cells. b KEGG pathway analysis of the hyperactivated kinases in ESR1 Y537S cells. Analysis was performed using the DAVID Bioinformatics Functional Annotation Tool. c Y537S/WT quantitative ratio of kinases in KiP analysis categorised by RTKs, PI3K, and MAPK pathways. Red bars represent Y537S-unique activated kinases. d MCF-7 cells were cultured in charcoal-stripped serum-supplemented media and analysed by immunoblot for phosphorylated and total RON. Numbers below each protein represent normalised densitometry of each protein compared to GAPDH. e T47D cells were cultured in charcoal-stripped serum-supplemented media and analysed by immunoblot for phosphorylated and total RON. Numbers below each protein represent normalised densitometry of each protein compared to GAPDH. f MCF-7 cells were cultured in full serum-supplemented media and treated with BMS-777607/ASLAN002 (RONi) for 90 min and analysed by immunoblot for phosphorylated and total RON and IGF-1R. Numbers below each protein represent normalised densitometry of each protein compared to GAPDH. g MCF-7 cells were cultured in full serum-supplemented media and transfected with indicated siRNA. Immunoblot analysis for phosphorylated and total IGF-1R was performed two days post transfection. Numbers below each protein represent normalised densitometry of each protein compared to GAPDH.