Research Paper Volume 12, Issue 6 pp 5280—5299

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.