Fig. 5: GnRHa counteracts CTX-induced ER stress.

a Expression of proteins including GRP78, ATF4, XBP1, and CHOP in the UPR pathway was assessed by western blotting. KGN cells were treated with 750 or 1000 µg/mL CTX, with or without GnRHa, for 36 h. b, c KGN cells were treated with CTX or CTX plus GnRHa for 36 h. mRNA levels were assessed by RT-qPCR for genes encoding proteins involved in the UPR (GRP78, XBP1, ATF4, and CHOP), and for genes encoding secretion-related proteins (SEC12 and SEC16). Those mRNAs were upregulated by CTX and downregulated when co-treated with GnRHa. d Typical immunofluorescence staining indicated the localization of AMH within KGN cells. Yellow in the merged images indicates the colocalization of AMH and GRP78. Nuclei were stained with DAPI. Scale bars represent 100 µm. e TM was used for simulation of ER stress, and expressions of proteins in the UPR pathway were assessed by western blotting, the results were consistent with the CTX-treated cells. f TM-induced upregulation of UPR sensors was confirmed by RT-PCR for mRNA expression. One-way ANOVA was used for (b, c, and f), error bars show the standard error of the mean. *P < 0.05, **P < 0.001, and ***P < 0.0001.