Fig. 5.

HFM1 knockdown disturbed the directional organelles transport of intercellular bridge. a Immunofluorescence staining for γ-tubulin (green) and DDX4 (red, dash line) at E17.5, P1 and P3. The nucleus was stained by Hoechst (blue). Scale bars: 20 μm. b The γ-tubulin foci area of a germ cell in cysts was generated by ImageJ software at E17.5 (n = 26 for WT and n = 21 for KO) and P1(n = 27 for WT and n = 20 for KO) (unpaired Student’s t-test, * P = 0.021, *** P < 0.001 vs. WT). c Timeline showing the 5-day culture scheme. d Fluorescence microscope analyses of adenovirus infection efficiency. Strong green fluorescence was observed under the fluorescent microscope following 5 days of adenovirus infection. Scale bars: 100px. e Western blotting analysis of HFM1 knockdown efficiency. HFM1 protein expression was down-regulated in Ad-shRNA-Hfm1 transfected ovaries after 5 days of culture (unpaired Student’s t-test, n = 3 independent experiments, * P = 0.014 vs. Control). f After 5 days of culture, ovaries transfected with Ad‐shRNA-Hfm1 stained with GM130 (red, white triangle) marking Golgi. DDX4 (green) is a germ cell marker. Hoechst (blue) was used to identify the nuclear DNA. Scale bars: 10 μm. g E17.5 ovaries were cultured with or without Ad‐shRNA-Hfm1 for 5 days, staining with ATP5A1 (green) and germ cell marker DDX4 (Red). Hoechst (blue) was used to identify the nuclear DNA. Scale bars: 20 μm. h The ATP5A1 foci area of a single germ cell was measured by ImageJ software (unpaired Student’s t-test, n = 30, *** P < 0.001 vs. Control). Data are presented as mean ± SEM