Fig. 1

Analysis of mRNA expression by semiquantitative RT-PCR/Southern blot. Total RNA was extracted from bovine GC collected from 2 to 4 mm small follicles (SF), dominant follicles (DF) at Day 5 of the estrous cycle, ovulatory follicles (OF) collected 24 h after injection of hCG, and corpora lutea (CL) from Day 5 of the estrous cycle, then used in mRNA expression analyses using semiquantitative RT-PCR/Southern blot analysis. GAPDH was used as a control gene and showed no significant difference in expression between groups. Gene-specific signals were normalized with corresponding GAPDH signals (1.8 kb) for each sample, and relative values are reported as percent of expression detected in OF. a) Expression of PLAT (2.5 kb) mRNAs was upregulated by 32.8-fold in OF compared with DF (P < 0.0001); expression of EGR1 mRNA (2.4 kb) was 24.7-fold higher in OF than in DF (P < 0.0001); expression of TNFAIP6 mRNA (1.6 kb) was upregulated by 7.9-fold in OF compared with DF (P < 0.0001); and expression of RGS2 mRNA (1.8 kb) was induced in OF compared with DF (P < 0.0001); b) expression of SAT1 (2.5 kb) mRNAs was upregulated by 18-fold in OF compared with DF (P < 0.0001); expression of GFPT2 mRNA (3 kb) was 26.8-fold higher in OF than in DF (P < 0.0029); expression of KIT mRNA (5 kb) was upregulated by 23.6-fold in OF compared with DF (P < 0.0001); and expression of POSTN mRNA (3.7 kb) was induced in OF compared with DF (P < 0.0001). Probability values for each one-way ANOVA analysis are specified above in parentheses. Different letters denote samples that differed significantly (P < 0.05) between group means for a specific gene. Data are presented as least-square means ± SEM, and the number of independent samples per group is indicated in parentheses