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Fig. 4 | BMC Biology

Fig. 4

From: CDC42 controls the activation of primordial follicles by regulating PI3K signaling in mouse oocytes

Fig. 4

CDC42 regulates PI3K signaling by binding to p110β in activated oocytes. a Ovaries at 3 dpp were stained for p110β (green) or CDC42 (red). Nuclei were dyed with a Hoechst (blue). Co-localization of p110β and CDC42 expression was observed in dormant oocytes (arrowheads) and on the intracellular membrane of activated oocytes (arrows). b Western blot showed that the expression of p110β increased in 5 and 7 dpp ovaries. c The interaction between CDC42 and p110β was detected in ovaries by Co-IP. The protein was extracted from 70 ovaries at 4 dpp. Approximately 5% of lysate was loaded as input. d The relationship between the activity of CDC42 and the interaction of CDC42 with p110β. The binding efficiency of CDC42 and p110β was significantly weakened in cultured 2 dpp ovaries after ML141 treatment. e Suppressing the activity of CDC42 with ML141 inhibited the intracellular localization of p110β in oocytes. A normal localization of CDC42 and p110β was observed in controls (arrows), while a cytoplasm localization of p110β was found in ML141 treated ovaries (arrowheads). f Ovaries at 1 dpp were transfected CDC42-DN or CDC42-CA lentiviral constructs for 2 days; empty lentivirus was used as control. Western blot showed that CDC42 expression was upregulated in ovaries after CDC42-DN or CDC42-CA transfections. g Quantification of ovarian follicles showed a decrease of activation (235.0 ± 53.9) in CDC42-DN ovaries and an increased activation (2245.0 ± 361.0) in CDC42-CA ovaries compared to controls (785.8 ± 93.5). h The interaction between CDC42 and p110β was examined in CDC42-DN and CDC42-CA treatment by Co-IP. The binding efficiency of CDC42 and p110β was significantly weakened in the CDC42-DN group but improved in the CDC42-CA group. The experiments were repeated at least three times, and representative images are shown. Scale bars, 50 μm

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