Supplementary Figure S12 from Autocrine 17-β-estradiol/estrogen receptor-α loop determines the response to immune-checkpoint inhibitors in non-small cell lung cancer

Dose-dependent inhibition of Akt, ERK1/2 and EGFR. ERα highNCI-H1975 cells were grown 24 h in fresh medium (Ctrl) or in medium containing 10 nM, 100 nM, 1 μM of the Akt inhibitor MK-2206 (Akti, panel a), 0.1 μM, 1 μM, 10 μM of the ERK1/2 inhibitor U-0126 (ERKi, panel b), 10 nM, 100 nM, 1 μM of the EGFR inhibitor AZD9291/Osimertinib (EGFRi, panel c). As read-out assays, Akt and ERK1/2 activity were measured by ELISA (technical duplicates), EGFR kinase activity was measured by a chemiluminescence-based assay (technical duplicates). Data are means + SD (n=3, biological replicates). *p<0.05, ***p<0.001: treated cells versus Ctrl cells (ANOVA).