Novel mutations in the L visual pigment gene found in Japanese men with protan color-vision defect having a normal order L/M gene array

Novel mutations in the L visual pigment gene found in Japanese men with protan color-vision defect having a normal order L/M gene arra

No significant difference in the differentiation and proliferation states between control and afadin cKO mice.

<p>Back skins from embryos at E14.5 were fixed and immunostained with the indicated Abs. Arrowheads in (<b>B</b>) indicate the cells double positive for Prox1 and BrdU. Scale bars in (<b>A</b>) and (<b>B</b>) represent 100 µm and 50 µm, respectively. Bar graphs show the number of Prox1-positive cells in the specific area of lymphatic vessels determined by podoplanin staining (<b>A</b>) and the percentage of BrdU-positive cells among Prox1-positive cells (<b>B</b>). Error bars indicate the mean ±S.D. from at least three independent experiments. N.S., not statistically significant.</p

Genotype and number of viable progeny from crosses between afadin^flox/+;Tie2-Cre and afadin^flox/flox mice.

<p>The percentage of each genotype relative to the total progeny at each embryonic and post natal day is shown in parentheses (). The number of mice with an abnormal phenotype is shown in brackets [ ].</p

Enhanced activation of RhoA in LMVECs by inactivation of afadin.

<p>(<b>A</b>) Activation of RhoA in BMVECs and LMVECs. At 2 days after transfection of siRNA or scramble RNA, BMVECs and LMVECs were lysed and then used for the pull-down assay, followed by western blotting with an anti-RhoA pAb. The numbers above the blot represent the relative density of GTP-bound RhoA normalized to the total amount of RhoA by comparing the value of BMVECs transfected with scramble RNA, which is expressed as 1.00. (<b>B, C</b>) Restoration of cell morphology and VE-cadherin-mediated cell-cell junctions by introduction of dominant-negative RhoA in afadin-knockdown LMVECs. LMVECs transfected with the indicated combination of siRNA, scramble RNA, pEGFP and pEGFP-RhoA-N19 were labeled with rhodamine-phalloidin to visualize F-actin (<b>B</b>) and with an anti-VE-cadherin mAb (<b>C</b>). Arrowheads indicate enhanced F-actin assembly (<b>B</b>) and reduced VE-cadherin staining (<b>C</b>). (<b>D</b>) VE-cadherin-mediated cell-cell junctions in control and afadin-knockdown BMVECs. BMVECs transfected with scramble RNA + pEGFP or siRNA + pEGFP were immunostained with an anti-VE-cadherin mAb. Bar graphs show the percentage of cells with VE-cadherin staining at cell-cell junctions. Bar graphs in <b><i>B</i></b>, <b><i>C</i></b> and <b><i>D</i></b> show the percentage of cells with enhanced F-actin assembly (<b>B</b>) and with VE-cadherin staining at cell-cell junctions (<b>C</b>) and (<b>D</b>). Error bars indicate the mean ±S.D. from at least three independent experiments. *, p<0.01 vs. Scramble +EGFP, and †, p<0.01 vs. Afadin siRNA +EGFP. N.S., not statistically significant. Scale bars represent 20 µm.</p

Disruption of cell-cell junctions on lymphatic, but not blood, vessel walls in afadin cKO mice.

<p>Back skins from control and afadin cKO embryos at E14.5 were fixed and immunostained with the indicated combination of Abs. Magnified images of the dashed boxes in (<b>B</b>) and (<b>E</b>) are also depicted. Arrows in (<b>A</b>) and (<b>D</b>) indicate the afadin-negative and VE-cadherin-positive area with patchy staining of VE-cadherin (<b>A</b>) and the afadin-negative and podoplanin-positive area with abnormal staining of podoplanin (<b>D</b>). White and yellow arrowheads in (<b>C</b>) indicate the arterial and venous vasculatures, respectively. Arrowheads in (<b>E</b>) indicate numerous punctures on lymphatic vessel walls. Bar graphs in (<b>B</b>) and (<b>E</b>) show the blood vessel density determined by VE-cadherin-positive and podoplanin-negative staining in the randomly selected areas (<b>B, left</b>), the percentage of VE-cadherin-positive staining in the podoplanin-positive areas (<b>B, right</b>) and the number of punctures in the LYVE-1-positive areas (<b>E</b>). Error bars indicate the mean ±S.D. at least three different samples. *, p<0.01 vs. Control. N.S., not statistically significant. Scale bars in (<b>A</b>), (<b>B</b>), (<b>C</b>), (<b>D</b>) and (<b>E</b>) represent 100 µm, 200 µm, 20 µm, 20 µm and 200 µm, respectively.</p