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. 2014 Jul 1;9(7):e101222. doi: 10.1371/journal.pone.0101222

Figure 1. Polyadenylation of cyclin B1 mRNA and a comparison of cyclin B1 and CPEB1 expression during IVM of porcine oocytes.

Figure 1

(A) Polyadenylation of cyclin B1 long (L-3′UTR) and short (S-3′UTR) mRNAs was examined by poly(A)-test in oocytes collected in different times of in vitro maturation (IVM). Corresponding maturation phases are figured below. The polyadenylation is highlighted by white lines next to each lane. (B) Oocytes collected after 0 to 44 h of IVM were subjected to western blot analysis of CPEB1 and cyclin B1 using specific antibodies. Detection of β-tubulin was used as a loading control. The phosphorylated form of CPEB1 is marked (*). (C) The protein expression of CPEB1 from three independent experiments was quantified using Quantity one software. The density of individual band was normalized to the total density of examined bands and the values of CPEB1 and phosphorylated CPEB1 were summed. The values represent the means ± SEM. (D) The protein expression of cyclin B1 from two independent experiments was quantified using Quantity one software. The density of individual band was normalized to the total density of examined bands. The values represent the means ± SEM. (E) Oocytes collected after 14, 16, 18 and 20 h of IVM were subjected to western blot analysis of CPEB1 and cyclin B1 using specific antibodies. Detection of GAPDH was used as a loading control.