Research Paper Volume 16, Issue 4 pp 3302—3331

Figure 6. ADSC-derived Exos modified by miR-204 inhibits METTL7A-medicated m6A methylation in HG-induced HK-2 cells. (A) m6A methylation level in HK-2 cells. (B) The expression of METTL3, METTL14, and METTL7A in HK-2 cells using western blotting. (A, B) HK-2 cells were exposed to HG (30 mM) and cultured for 24 h to induce an in vitro model of DN. To figure out the impact of exosomal miR-204 in m6A methylation in DN, ADSCs were transfected with miR-204 mimic or miR NC (negative control) for 48 h. Next, Exos were isolated from the transfected ADSCs and co-cultured with HK-2 a for 12 h at the concentration of 100 μg/mL. Data were expressed as mean ± standard deviation. ^**p < 0.01 vs. Control group; ^##p < 0.01 vs. HG group; ^&&p < 0.01 vs. HG + Exo-miR NC group; ns indicates no significant differences between groups. (C, D) The expression of miR-204 and METTL7A in HEK293T cells using qRT-PCR and western blotting. (C, D) Data were expressed as mean ± standard deviation. ^**p < 0.01 vs. mimic NC group; ^{^^}p < 0.01 vs. inhibitor NC group. (E) Determination of the interaction between miR-204 and METTL7A in HEK293T cells using dual-luciferase reporter assay. Data were expressed as mean ± standard deviation. ^**p < 0.01 vs. miR NC group; ns indicates no significant differences between groups. Abbreviations: ADSC: adipose-derived stem cell; Exos: exosomes; miR-204: microRNA-204; METTL: methyltransferase-like; m6A: N6-methyladenosine; HG: high glucose; qRT-PCR: quantitative real-time polymerase chain reaction.