Figure 3: miR-125b is repressed by an NSun2-mediated methylation in gonadotrope cells.
From: A regulatory loop between miR-132 and miR-125b involved in gonadotrope cells desensitization to GnRH
(a,b) Effect of overexpressing or blocking NSun2. LβT2 cells were electroporated with NSun2 or control expressing vector and miR-125b, Fshb and Lhb mRNA levels were quantified by qRT-PCR and normalized to snU6 (miR-125b) or to Gapdh. (a) Overexpression of NSun2 reduced mi-125b and increased Fshb mRNA level. Lhb mRNA level was not affected (n = 9). (b) Blocking NSun2 using an anti-NSun2 LNA led to inverse effects compared to cells electroporated with a scramble LNA (n = 9). (c) Effect of a GnRHa treatment and of overexpressing NSun2 on the methylation status of miR-125b. LβT2 cells were either treated by GnRH (10 nM for 4 h) or electroporated with an NSun2 expressing vector and harvested. Total RNA extract was immunoprecipitated using an anti-m6A antibody and miR-125b was quantified in the immunoprecipitated fraction and normalized to input miR-125b level. Overexpression of NSun2 had a similar stimulatory effect as the GnRHa treatment on the methylation status of miR-125b (n ≥ 4). *P < 0.05; **P < 0.01; ***P < 0.001.
