Figure 6.

Oocyte-specific knockout of Lonp1 leads to AIFM1 nuclear translocation.

(a) LONP1 colocalized with AIFM1 in wild-type mouse oocytes, as indicated by immunofluorescence analysis (at least 30 GV oocytes were included). (b) Immunoprecipitation of LONP1 and AIFM1 in mouse oocytes (approximately 5000 wild-type GV oocytes were used). (c) FPKM values of Capn1 in oocytes from secondary follicles of Lonp1^loxp/loxp and Lonp1^Gdf9Cre cKO mice (n=3 for each group). (d) qRT-PCR analysis of Capn1 in oocytes from secondary follicles of Lonp1^loxp/loxp and Lonp1^Gdf9Cre cKO mice (n=9 for each group). (e) Immunofluorescence staining of CALPAIN1 in oocytes from secondary follicles of Lonp1^loxp/loxp and Lonp1^Gdf9Cre cKO mice. Scale bar = 50 μm. (f and g) Immunofluorescence staining of AIFM1 in the ovaries of 3-week-old Lonp1^loxp/loxp and Lonp1^Gdf9Cre cKO mice (n=3 for each genotype). The asterisks indicate nuclei (*p < 0.05, Student's t test). Scale bar = 50 μm. (h) Blastocyst formation in oocytes from 3-week-old Lonp1^GdfCre cKO mice after pretreatment with the PARP inhibitor 3-aminobenzamide to rescue AIFM1-induced apoptosis (n>30). (i) Immunofluorescence staining of AIFM1 in 293T cells after Lonp1 knockdown. Scale bar = 20 μm. Scale bar = 50 μm. (j and k) Immunoprecipitation of LONP1 and AIFM1 from the cytoplasmic and nuclear proteins of 293T cells after Lonp1 knockdown.