Fig. 6

Irisin mediates skeletal muscle-ovary crosstalk and inhibits the IRE1α-TXNIP/ROS-NLRP3 signaling pathway in TCs. TCs were treated with DHT, in the presence or absence various concentrations of irisin. A Cell viability was measured by CCK8. B and D The protein (B) and mRNA (D) levels of IRE1α, TXNIP and NLRP3 in TCs were analyzed by western blot and qRT-PCR. (C) The expression of NLRP3 in TCs was analyzed by immunofluorescence staining (60 ×). E–F TC MDA levels (E) and SOD activity (F) were analyzed using an enzymatic colorimetric method. G ROS in TCs 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