Fig. 3. Kras^G12D conditional knock-in mice develop ovarian lesions with altered granulosa cell proliferation, differentiation and apoptosis. (A) Size differences in wild-type (WT) and LSL-Kras^G12D;Cyp19-Cre ovaries at various ages. (B-D) Histology of WT (B) and LSL-Kras^G12D;Cyp19-Cre (C,D) ovaries at 6 months of age. NF, normal follicle; AF, abnormal follicle. (E,F) BrdU incorporation assay in 12-week-old WT (E) and LSL-Kras^G12D;Cyp19-Cre (F) ovaries. Abnormal follicle-like structures are indicated by arrows (as below). (G,H) Immunofluorescent detection of phospho-histone H3 (pHH3, red) in 12-week-old WT (G) and LSL-Kras^G12D;Cyp19-Cre (H) ovaries. (I,J) BrdU incorporation (I) and immunofluorescence for the mitosis marker phospho-histone H3 (J) indicate slightly increased levels of proliferation in Kras^G12D-expressing granulosa cells of antral follicles, as compared with wild type. (K,L) Immunohistochemical detection of PCNA in 12-week-old WT (K) and LSL-Kras^G12D;Cyp19-Cre (L) ovaries. (M,N) Apoptosis assays in 4-week-old WT (M) and LSL-Kras^G12D;Cyp19-Cre (N) ovaries, 2 hours after hCG treatment. (O,P) Immunofluorescent detection of cleaved caspase 3 (CC3) in 12-week-old WT (O) and LSL-Kras^G12D;Cyp19-Cre (P) ovaries.