FIG. 2.

Treatment with oxamflatin increased the acetylation levels of α-tubulin during the first and second mitotic cell cycle of porcine SCNT embryos through downregulating HDAC6. (A and B) The relative expression levels of HDAC6 at two-cell- (24–28 h, postactivation) and four-cell- (48–52 h, postactivation) stage embryos between C-NT and T-NT groups, respectively. The relative expression level of mRNA was normalized with β-actin. (C) Acetylation levels of α-tubulin at the first cell cycle after treatment with or without oxamflatin. Arrows indicate the second meiotic midbody at prophase, spindle at anaphase, and the midbody at telophase, respectively. (D) Acetylation levels of α-tubulin at the second cell cycle after treatment with or without oxamflatin. Arrows indicate the spindle at metaphase and the midbody at telophase, respectively. The experiments were repeated three times and each replication included at least 10–15 embryos. Representative examples are shown. Scale bar, 20 μm. (E and F) Quantification of mean fluorescence signal intensities of Ac-α-tubulin at the first cell cycle (prophase, anaphase, telophase) or the second cell cycle (metaphase, telophase) of porcine SCNT embryos between C-NT and T-NT groups. T-NT, SCNT embryos treated with 1 μM oxamflatin for 15 h postactivation; C-NT, nontreated SCNT embryos. Data are presented as mean±standard deviation (SD). (*) p<0.05; (**) p<0.01.