Fig 2: miR-144-3p attenuates cisplatin-induced primordial follicle activation via MAP3K9 in mouse ovaries in vivo. A Western blotting detection of MAP3K9 and PI3K/AKT pathway-related proteins in mice. B H&E staining displaying ovarian morphology and columns representing the proportion of follicles at various stages of development in mice. Control group, n = 3; POF + agomir NC group, n = 3; POF + agomir group, n = 3. H&E: hematoxylin and erosion; POF: premature ovarian failure. *P < 0.05; **P < 0.01, two-tailed Student’s t test

Fig 3: Map3k9 is upregulated in GCs of POF mice. A Bioinformatics prediction combined with mRNA microarray analysis were performed to identify the potential target genes of miR-144-3p. B Differentially expressed gene pathway enrichment analysis in the POF group and the control group. C Real-time PCR was used to evaluate the relative expression of target genes of miR-144-3p in the POF group and the control group. D Protein levels of MAP3K9 in both groups were detected using western blotting. E Immunohistochemistry analysis of the protein expression of MAP3K9 in both groups. Control group, n = 10; POF group, n = 10. GCs: granulosa cells; POF: premature ovarian failure. *P < 0.05; **P < 0.01, two-tailed Student’s t test. Scale bar, 100 μm

Fig 4: Effects of miR-204-3p on apoptosis-related proteins and necroptosis-related proteins in AGS and HGC-27 cells. A and B: Effect of miR-204-3p on apoptosis-related proteins BCL-2, BAX and Caspase-3 in AGS cells; C and D: Effect of miR-204-3p on apoptosis-related proteins BCL-2, BAX and Caspase-3 in HGC-27 cells; E and F: Effects of miR-204-3p on necroptosis-related proteins RIP1, p-RIP1, MLK1, and p-MLK1 in AGS cells; G and H: Effects of miR-204-3p on necroptosis-related proteins RIP1, p-RIP1, MLK1, and p-MLK1 in HGC-27 cells. aP < 0.05, bP < 0.01, cP < 0.001. NC: Empty lentiviral vector; KD: miR-204-3p knockdown lentiviral vector; OE: miR-204-3p overexpression lentiviral vector.

Fig 5: miR-144-3p protects GCs against cisplatin-induced apoptosis by targeting MAP3K9 via the p38 MAPK pathway. A Western blotting to detect the protein levels of cleaved caspase-3 and p38 MAPK pathway-related molecules in cisplatin injured COV434 cells. B Western blotting to determine the level of MAP3K9 in COV434 cells transfected with MAP3K9 siRNA and scrambled control oligonucleotide. C Cleaved caspase-3 and p38 MAPK pathway-related proteins levels were analyzed by western blotting in miR-144-3p antagomir and MAP3K9 siRNA co-treated COV434 cells. GCs, granulosa cells. *P < 0.05; **P < 0.01, two-tailed Student’s t test