Fig. 5

Irisin mediates skeletal muscle-ovary crosstalk and inhibits the IRE1α-TXNIP/ROS-NLRP3 signaling pathway in GCs. GCs were treated with DHT, in the presence or absence of various concentrations of irisin. A-B Cell viability was measured by CCK8. (C-D) The mRNA (C) and protein (D) levels of IRE1α, TXNIP and NLRP3 in GCs were analyzed by qRT-PCR and western blot. (E) The expression of IRE1α in GCs was analyzed by immunofluorescence staining (60 ×). F-G GC MDA levels (F) and SOD activity (G) were analyzed using an enzymatic colorimetric method. H ROS in GCs was assessed by the DCF-DA probe using confocal microscopy (90 ×). Data are shown as the mean ± SD. *p ≤ 0.05, **p ≤ 0.01. DHT, dihydrotestosterone; IRE1α, inositol-requiring enzyme 1α; TXNIP, thioredoxin-interacting protein; NLRP3, NOD-like receptor family pyrin domain containing 3; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Ctrl, control; DAPI, 4’,6-diamidino-2-phenylindole; MDA, malondialdehyde; SOD, superoxide dismutase; DCF-DA, dichlorofluorescein diacetate; CCK8, Cell Counting Kit-8