Similarity between bovine SOLD1 and the Ly-6 domain of related molecules.

<p>Similarity between bovine SOLD1 and the Ly-6 domain of related molecules.</p

Expression of <i>SOLD1</i> mRNA.

<p>(A) Expression of <i>SOLD1</i> mRNA in various bovine tissues, including heart, liver, lung, spleen, and kidney, was analyzed by RT-PCR. Cotyledonary tissue at Day 150 of gestation was used as a bovine placental sample. <i>GAPDH</i> expression in each tissue is presented as a positive control.<i>CSH1</i> expression in each tissue except for the placenta is presented as a negative control (NC). <i>ALB</i> expression in the other tissue is presented as a negative control (NC).(B)Quantitative expression of SOLD1 in bovine placental tissues during the initial to rate stage of pregnancy by qRT-PCR analysis. Pre-Im, pre-implantaion; Peri-Im, peri-implantation; and Post-Im, post-implantation. CON, conceptus; EEM, extra-embryonic membrane; EMB, embryo; COT, cotyledon; and ICOT, intercotyledon. D, day after insemination. Expression of these mRNAs was normalized to the expression of <i>GAPDH</i> measured in the corresponding RNA preparation. Values are means±SEM. Values with different letters (a, b, c and d) are significantly different (P<0.05).</p

Recombinant SOLD1 binding assay to ECM-coated plate.

<p>(A) Absorbance of SOLD1 binding to 96-well microtiter plates coated commercially with ECM components type I collagen (COL1), type IV collagen (COL4), fibronectin (FN), and laminin (LAM), and the negative control, poly-L-lysine (PLL). (B) Absorbance of SOLD1 binding to 96-well microtiter plates coated manually with ECM components (300 µg/well each) telopeptide including type I collagen (tropocollagen, COL1-A), telopeptide excluding type I collagen (atelocollagen, COL1-P), type III collagen (COL3), and type IV collagen (COL4). The means±SEM of triplicate measurements are shown. Values with different letters are significantly different (P<0.05).</p

mRNA localization of <i>SOLD1</i> in the bovine placentome on Day 60 of gestation.

<p><i>SOLD1</i> mRNA was detected by in situ hybridization. (A, D) DIG-labeled anti-sense cRNA probes were used. (B, E) Enlarged images of frames in A and D, respectively. (C, F) DIG-labeled sense cRNA probes were used. CE, caruncular epithelium; CS, caruncular stroma; T, trophoblast; TMC, trophoblast mononucleate cells; BNC, trophoblast binucleate cells; MPV, mesenchyme of primary villi; and MSV, mesenchyme of secondary villi. Scale bars = 100 µm (A, C, D and F) and 20 µm (B and E).</p

Protein localization of COL1 and COL3 in the bovine placentome on Day 60 of gestation.

<p>(A) COL1 protein was detected in the bovine placentome. (B) COL3 protein was detected in the bovine placentome. CE, carunclar epithelium; CS, carunclar stroma; T, trophoblast; MPV, mesenchyme of primary villi; and MSV, mesenchyme of secondary villi. Scale bars = 100 µm.</p

Comparison of the Ly-6 domain sequence and the genome structure of <i>SOLD1</i>.

<p>(A) Comparison of amino acid sequences between ruminant SOLD1 and the phylogenetically neighbouring Ly-6 domains in each protein selected by ProDom. Residues identical in all Ly-6 domains are shown by blue letters. Sequence gaps are shown by hyphens.The potential <i>N</i>-glycosylation site is underlined in purple.(B) The genome structure of the <i>SOLD1</i> region (genome sequence position 29,756,350 to 29,757,780) in bovine chromosome 17, as determined by NCBI MapView.</p

Oligonucleotide primers used for cDNA cloning or RT-PCR analysis.

*<p>Position: The nucleotide position in each accession number.</p

Apico-basolateral polarity of SOLD1 secretion from BT-1 cells.

<p>(A) Western blot analysis of BT-1 conditioned medium and BT-1 cell lysate. (B) Results of the SOLD1 binding assay. BT-1 cells were cultured and removed on COL1-A coated plates. The plates were washed ten times with TBST. SOLD1 binding to COL1-A coated plates was measured by monitoring absorbance at 405 nm in each of four wells per sample. The means±SEM of triplicate measurements are shown. Values with asterisks are significantly different (P<0.05).</p

Oligonucleotide primers used for qRT-PCR analysis.

*<p>Position: The nucleotide position in each accession number</p

Protein localization of SOLD1 in the bovine placentome on Day 60 of gestation.

<p>(A, C) SOLD1 protein was detected by immunohistochemistry. (B, D) Enlarged images of frames in A and C, respectively. Custom-made bovine anti-SOLD1 antibody was used. CE, carunclar epithelium; CS, carunclar stroma; T, trophoblast; TMC, trophoblast mononucleate cells; BNC, trophoblast binucleate cells; MPV, mesenchyme of primary villi; and MSV, mesenchyme of secondary villi. Scale bars = 100 µm (A and C) and 20 µm (B and D).</p