AlkB Homolog H5 Attenuates TGF-β1-Induced Epithelial-Mesenchymal Transition of Retinal Pigment Epithelial Cells via Regulating Zinc Finger E-box Binding Homeobox 1
AlkB Homolog H5 Attenuates TGF-β1-Induced Epithelial-Mesenchymal Transition of Retinal Pigment Epithelial Cells via Regulating Zinc Finger E-box Binding Homeobox 1
Yunzhi Lin1, Jing He1, Xingwei Wu1* and Qian Shi2*
TGF-β1 induces ARPE-19 EMT. (A), CCK-8 assay examined ARPE-19 cell viability in TGF-β1 and control groups; (B), Transwell assay examined ARPE-19 cell migration in TGF-β1 and control groups; (C), EMT markers expressions were unclosed by RT-qPCR in TGF-β1 and control groups.
ALKBH5 suppresses ZEB1 mRNA stability in an m6A-IGF2BP1-dependent manner. (A), RT-qPCR determined ZEB1 expression after depletion of ALKBH5 in untreated ARPE-19 cells and after ALKBH5 overexpression in TGF-β1-treated ARPE-19 cells; (B), Luciferase reporter assay unclosed the binding of ALKBH5 with ZEB1 promoter in ARPE-19 cells; (C), The stability of ZEB1 mRNA was determined by RT-qPCR after depletion or overexpression of ALKBH5 in ARPE-19 cells treated with α-amanitin; (D), MeRIP examined m6A modification of ZEB1 after depletion or overexpression of ALKBH5 in ARPE-19 cells; (E), RT-qPCR demonstrated the interference efficiency (sh-NC, sh-IGF2BP1#1 and sh-IGF2BP1#2) and the overexpression efficiency (Vector and IGF2BP1) in ARPE-19 cells; (F), RT-qPCR examined ZEB1 expression after depletion or overexpression of IGF2BP1 in ARPE-19 cells; (G), RIP assay validated the interaction between IGF2BP1 and ZEB1 in ARPE-19 cells; (H), RT-qPCR unclosed IGF2BP1 and ZEB1 expressions in ARPE-19 and TGF-β1-treated ARPE-19 cells under indicated transfections.
ALKBH5 modulates ZEB1 to affect EMT of ARPE-19 cells. (A), CCK-8 assay examined cell viability in ARPE-19 cells transfected with sh-NC, sh-ALKBH5#1 or sh-ALKBH5#1+ZEB1 and in TGF-β1-treated ARPE-19 cells transfected with Vector, ALKBH5 or ALKBH5+sh-ZEB1#1; (B), Transwell assay examined ARPE-19 cell migration in different transfection groups; (C), EMT markers expressions were uncovered by RT-qPCR in different transfection groups.
