Fig. 1: E2 activated mTOR-mediated m6A modification.
From: mTORC1 accelerates osteosarcoma progression via m6A-dependent stabilization of USP7 mRNA
A, B The protein expression levels of p-S6, S6, p-S6K, and S6K in MG63 and U2OS cells after treatment with 0–1000 nM of E2 for 48 h. C The protein expression levels of MTTLE3, METTL14, and WTAP in U2OS cells after treatment with or without 100 nM of E2 for 48 h. D The protein expression levels of MTTLE3, METTL14, and WTAP in U2OS cells after treatment with 100 nM of E2 for 48 h with or without pretreatment of 1 μM of torin1. E LC–MS analysis of m6A levels in U2OS cells treated with with 100 nM of E2 for 48 h with or without pretreatment of 1 μM of torin1. F, G Determination and quantification of ASC speck formation by immunofluorescence staining in 100 nM of E2 treated U2OS cells with or without torin1, scale bar: 50 µm. Arrows signify ASC speck. H The mRNA expression levels were determined following following shCtrl, shWTAP#1, and shWTAP#2 transfection. I The relative cell proliferation in shCtrl, shWTAP#1, and shWTAP#2 transfected U2OS cells for 0–96 h. The EdU staining (J) and EdU positive cells (K) in shCtrl, shWTAP#1, and shWTAP#2 transfected U2OS cells for 48 h, scale bar: 50 µm. L, M The migration capacity and migrated cell number in shCtrl, shWTAP#1, and shWTAP#2 transfected U2OS cells for 24 h, scale bar: 50 µm. N LC–MS analysis of m6A levels in U2OS cells treated with with 100 nM of E2 following shCtrl or shWTAP#2 transfection. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 vs control group, ##P < 0.01, ####P < 0.0001 vs E2 group.
