Proliferation of cultured LSCs.

<p>(A) Cellular proliferation was determined by DNA assay at the indicated times with or without LH (10 ng/ml) after the LSCs were seeded (n=5). Daily observation of luteal steroidogenic cells at (B) 1, (C) 4 days from developing CL and (D) 1, (E) 4 days from mid CL were shown. To confirm that cultured cells were steroidogenic cells, the LSCs isolated from mid luteal stage were cultured for 24 h, then immunohistochemical staining with HSD3B was performed (F). PBS was used as a negative control (G). (H) and (I) show the effect of LH (10 ng/ml) on cellular proliferation of bovine luteal cells isolated from the developing and mid CL, respectively (mean±SEM, n-5 experiments). <i>Asterisks</i> indicate that LH had a significant effect on DNA concentration (P<0.05), as determined by ANOVA followed by Fisher’s PLSD as a multiple comparison test.</p

KI-67 and HSD3B expression in the bovine CL.

<p>Representative images of double-immunohistochemical staining for KI-67 and HSD3B in early (A), developing (B) and mid (C) bovine CL. Normal horse serum was used as a negative control (D). Scale bars represent 50 μm. (E) Percentage of KI-67 positive LSCs in the bovine CL (mean±SEM, n=3 experiments). Percentages in each luteal stage were derived from dividing the number of Ki-67 and HSD3B positive cells by total number of HSD3B positive cells. Cells were counted in three randomly chosen areas. Different superscript letters indicate significant differences (P<0.05).</p

Effect of LH on cell cycle-related genes and <i>PTEN</i> expression in cultured LSCs.

<p><i>CCND2</i>, <i>CCNE1</i>, <i>CDKN1A</i>, <i>CDKN1B</i> and <i>PTEN</i> mRNA in cultured LSCs isolated from the developing (A) and mid (B) CL (mean±SEM, n=4 experiments). The cells were exposed to LH (10 ng/ml) for 24 h. Asterisks indicate significant differences (P<0.05), as determined by ANOVA followed by Fisher’s PLSD as a multiple comparison test.</p

KI-67 expression in the bovine CL.

<p>Representative images of immunohistochemical staining for KI-67 in the bovine CL throughout the estrous cycle (A, B; early, C, D; developing, E, F; mid, G, H; late, I, J; regressed luteal stages). Scale bars represent 100 μm (low magnitude) and 50 μm (high magnitude).</p

Cell cycle-related genes and <i>PTEN</i> expression in bovine freshly isolated LSCs.

<p>Fold differences of cell cycle-related genes such as <i>CCND2</i>, <i>CCNE1</i>, <i>CDKN1A</i>, <i>CDKN1B</i> and tumor suppressor gene <i>PTEN</i> mRNA between freshly isolated developing LSCs and mid LSCs. Single and double <i>Asterisks</i> indicate significant differences (* ; P<0.1, ** ; P<0.05), as determined by ANOVA followed by Fisher’s PLSD as a multiple comparison test.</p

Additional file 1: of Differential gene expression profiling of endometrium during the mid-luteal phase of the estrous cycle between a repeat breeder (RB) and non-RB cows

Table S1. List of up- and down-regulated genes in CAR of ipsilateral uterine horns of RB cows (n = 4) on Day 15 of the estrous cycle as compared with non-RB cows (n = 4). Table S2. List of up- and down-regulated genes in ICAR of ipsilateral uterine horns of RB cows on Day 15 of the estrous cycle as compared with non-RB cows. Table S3. List of up- and down-regulated genes in CAR of contralateral uterine horns of RB cows on Day 15 of the estrous cycle as compared with non-RB cows. Table S4. List of up- and down-regulated genes in ICAR of contralateral uterine horns of RB cows on Day 15 of the estrous cycle as compared with non-RB cows. (XLSX 254 kb