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. 2022 Jan 27;13(1):87. doi: 10.1038/s41419-022-04541-1

Fig. 5. Effect of acute fasting on glycolysis and mouse primordial follicle activation in vivo.

Fig. 5

Two-day-old female mice were kept with their mother (control), or were separated from their mother for 18 h and then returned to their mother (acute fasting). The ovaries were collected at 24 h (c–f) and 48 h (a, b, g, h) of treatment, respectively. a, b Morphological comparison of the ovaries (a) and the number of primordial and growing follicles (arrows, b) in the control and acute fasting group. Nuclei were stained by hematoxylin. Scale bars: 50 μm. c The mRNA levels of Glut4, Hk1, Pfkl, Aldoa, Eno1, Tpi, Pkm2, and Ldhb in the control and acute fasting group. d The protein levels of GLUT4, HK1, PFKL, and PKM2 in the control and acute fasting group. e, f The protein levels of p-AMPK, p-mTOR (e), p-Akt, and p-FOXO3a (f) in the control and acute fasting group. g, h The localization of FOXO3a in oocyte nuclear (arrowheads) and cytoplasm (arrows, g) and the percentage of oocytes with FOXO3a nuclear export (h) in each section in the control and acute fasting group. FOXO3a, green; DDX4, red; DAPI, blue. Scale bars: 50 μm. Fasting, acute fasting. All the experiments were repeated three times, and the representative images are shown. In western blot results, the levels of total AMPK, mTOR, Akt, and FOXO3a were used as the corresponding internal control for p-AMPK, p-mTOR, p-Akt, and p-FOXO3a, respectively, and β-actin was used as the internal control for GLUT4, HK1, PFKL, and PKM2. Bars indicate the mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001 vs. control.