Figure 3.

Leptin inhibits the m⁶A methylation of Plin5 through FTO. (A) Porcine adipocytes in vitro were treated with 50 nmol/μL leptin for 24 h to detect the mRNA expression of methylation-related genes. (B) The protein expression of FTO under the same treatment as (A). (C) Dot Blot detected the level of total RNA m⁶A in 50 nmol/μL leptin-treated porcine adipocytes in vitro. (D) The colorimetric assay was used to quantify the relative m⁶A levels of 50 nmol/μL leptin-treated porcine adipocytes in vitro. (E) m⁶A-IP was used to detect Plin5 M⁶A methylation in 50 nmol/μL leptin-treated porcine adipocytes in vitro. (F) Real-time quantitative PCR was used to detect FTO overexpression and interference efficiency in vitro. (G) m⁶A-IP detected the m⁶A level of Plin5 after overexpression or interference with FTO in vitro. (H) The relative M⁶A level of 50 nmol/μL leptin treated porcine adipocytes in vitro after overexpression or interference with FTO. (I) Plin5 M⁶A levels in 50 nmol/μL leptin-treated porcine adipocytes in vitro after overexpression or interference with FTO. n = 6 for each group of cell samples, values are means ± SD. vs. control group, * p < 0.05, ** p < 0.01, *** p < 0.001.