Fig. 4

KIFC3 regulates stability of spindle microtubules (A) Control and KIFC3-antibody injection oocytes after cold treatment at the MI stage were stained with anti-α-tubulin (green) and counterstained with Hoechst 33342 to visualize the chromosomes (blue). Bar = 20 μm. (B) The rate of abnormal spindle morphology after cold treatment of KIFC3 antibody injection and control oocytes at the MI stage. Graph shows means ± sem of results obtained in 3 independent experiments. *P < 0.05. (C) Control and KIFC3-antibody injection oocytes after Nocodazole treatment at the MI stage were stained with anti-α-tubulin (green) and counterstained with Hoechst 33342 to visualize the chromosomes (blue). Bar = 20 μm. (D) The relative area of spindle after nocodazole treatment of KIFC3 antibody injection and control oocytes at the MI stage. Graph shows means ± sem of results obtained in 3 independent experiments. *P < 0.05. (E) Co-IP was performed with an anti-KIFC3 antibody. The immunoblots of protein precipitates were probed with anti-Sitr2 and anti- acetylated tubulin antibody. (F) Immunofluorescence analysis of acetylated tubulin (red) and α-tubulin (green) in control and KIFC3 antibody injection oocytes. Bar = 10 μm. (G) The histogram shows the relative fluorescence intensity of acetylated tubulin signals in control and KIFC3 antibody injection oocytes. Graph shows means ± sem of results obtained in 3 independent experiments. **P < 0.01. (H) Immunofluorescence analysis of Sirt2 (red) and α-tubulin (green) in control and KIFC3 antibody injection oocytes. Bar = 20 μm. (I) The histogram shows the relative fluorescence intensity of Sirt2 signals in control and KIFC3 antibody injection oocytes. Graph shows means ± sem of results obtained in 3 independent experiments. *P < 0.05