Fig. 5: P62 targets the OXPHOS signaling pathway to regulate LH and GnRH-induced LH production.
From: Pituitary P62 deficiency leads to female infertility by impairing luteinizing hormone production
Rescue experiments were conducted in LβT2 cells by silencing p62 with shRNA, and p62 was stably silenced in cells overexpressing Ndufa2 with a plasmid for 48 h, thus generating three groups of LβT2 cells: shRNA-con, p-con; shRNA-p62, p-con; shRNA-p62, p-Ndufa2. a RT-PCR detection of p62 and Ndufa2 to confirm transfection efficiency, n = 3. b, c mRNA detection of Lhb (n = 3) and measurement of supernatant LH (n = 6–8) in each group. d–g Seahorse assay analysis of mitochondrial respiratory function in LβT2 cells, including ATP production, basal respiration, and maximal respiration, in each group, n = 5. h Relative serum LH in p62flox/flox αGSUcre and p62flox/flox female mice treated with vehicle or the GnRH agonist gonadorelin (7.5 µg/kg body weight), n = 3-4. i Relative Lhb mRNA after gonadorelin induction in LβT2 cells, and j supernatant LH concentration level after treatment with the calcium chelator BAPTA (2 mM) or the mitochondrial NADH: ubiquinone reductase inhibitor rotenone (5 μM), with/without gonadorelin (100 nM), n = 4. All data are presented as the mean ± SD. Statistical analyses were performed using one-way ANOVA and Student’s t-test (b, i). *P < 0.05; **P < 0.01; ***P ≤ 0.001.
