Research Paper Volume 12, Issue 6 pp 5280—5299

m6A-induced lncRNA MALAT1 aggravates renal fibrogenesis in obstructive nephropathy through the miR-145/FAK pathway

Figure 8. mA modification participates in the upregulation of MALAT1 in renal fibrosis. (A) ELISA was used to measure the m6A levels in HK2 cells treated with TGF-β1 at different concentrations for approximately 48 h. (B and C) qPCR and western blot analyses of METTL3, METTL14 and WTAP in HK2 cells treated with TGF-β1 at different concentrations for approximately 48 h. (D) qPCR analyses of METTL3 expression in HK2 cells transfected with siMETTL3 or siNC for approximately 48 h. (E) qPCR analyses of E-cad and α-SMA expressions in HK2 cells treated with siNC, siNC+TGF-β1 and siMETTL3+TGF-β1 for approximately 48 h. (F and G) CCK8, EdU and cell migration analyses of the viability, proliferation and migration potential of HK2 cells treated with siNC, siNC+TGF-β1 and siMETTL3+TGF-β1 for approximately 48 h. (H) qPCR analyses of MALAT1 expression in HK2 cells transfected with siMETTL3 or siNC for approximately 48 h. (I) METTL3 RIP-qPCR analysis of MALAT1 in HK2 cells. (J) m6A RIP-qPCR analysis of MALAT1 in HK2 cells transfected with siMETTL3 or siNC for approximately 48 h. (K) the predicted m6A sites (with high confidence) in MALAT1. (L) the mutated m6A sites of MALAT1. (M) the inhibitory role of shMETTL3 was attenuated in the luciferase activity inserted the MALAT1 with mutant type (Mut) m6A sites. GAPDH was used as a control. *P < 0.05 and **P < 0.01.