Theoretical curves for the Fung, Gent and Ogden models at each loading velocity.

<p>Theoretical curves for the Fung, Gent and Ogden models at each loading velocity.</p

Repeatability of measurements of infant porcine cerebellum tissue up to 30% strain at loading velocity of 10 mms-1 (strain rate 2s-1),100 mms-1 (strain rate 20 s-1), and 500 mms-1 (strain rate100 s-1).

<p>Repeatability of measurements of infant porcine cerebellum tissue up to 30% strain at loading velocity of 10 mms-1 (strain rate 2s-1),100 mms-1 (strain rate 20 s-1), and 500 mms-1 (strain rate100 s-1).</p

Retinoic Acid Signaling Plays a Restrictive Role in Zebrafish Primitive Myelopoiesis

<div><p>Retinoic acid (RA) is known to regulate definitive myelopoiesis but its role in vertebrate primitive myelopoiesis remains unclear. Here we report that zebrafish primitive myelopoiesis is restricted by RA in a dose dependent manner mainly before 11 hpf (hours post fertilization) when anterior hemangioblasts are initiated to form. RA treatment significantly reduces expressions of anterior hemangioblast markers <em>scl</em>, <em>lmo2</em>, <em>gata2</em> and <em>etsrp</em> in the rostral end of ALPM (anterior lateral plate mesoderm) of the embryos. The result indicates that RA restricts primitive myelopoiesis by suppressing formation of anterior hemangioblasts. Analyses of ALPM formation suggest that the defective primitive myelopoiesis resulting from RA treatment before late gastrulation may be secondary to global loss of cells for ALPM fate whereas the developmental defect resulting from RA treatment during 10–11 hpf should be due to ALPM patterning shift. Overexpressions of <em>scl</em> and <em>lmo2</em> partially rescue the block of primitive myelopoiesis in the embryos treated with 250 nM RA during 10–11 hpf, suggesting RA acts upstream of <em>scl</em> to control primitive myelopoiesis. However, the RA treatment blocks the increased primitive myelopoiesis caused by overexpressing <em>gata4/6</em> whereas the abolished primitive myelopoiesis in <em>gata4/5/6</em> depleted embryos is well rescued by 4-diethylamino-benzaldehyde, a retinal dehydrogenase inhibitor, or partially rescued by knocking down <em>aldh1a2</em>, the major retinal dehydrogenase gene that is responsible for RA synthesis during early development. Consistently, overexpressing <em>gata4/6</em> inhibits <em>aldh1a2</em> expression whereas depleting <em>gata4/5/6</em> increases <em>aldh1a2</em> expression. The results reveal that RA signaling acts downstream of <em>gata4/5/6</em> to control primitive myelopoiesis. But, 4-diethylamino-benzaldehyde fails to rescue the defective primitive myelopoiesis in either <em>cloche</em> embryos or <em>lycat</em> morphants. Taken together, our results demonstrate that RA signaling restricts zebrafish primitive myelopoiesis through acting downstream of <em>gata4/5/6</em>, upstream of, or parallel to, <em>cloche</em>, and upstream of <em>scl</em>.</p> </div

Primers used to make constructs to analyse promoter activity in the study.

<p>C2-A, C2-B, C2-C, C2-D, C2-E, C2-a, C2-b, C2-c and C2-d: Forward primers for construction of the luciferase reporter gene vectors pGL₃-A, pGL₃-B, pGL₃-C, pGL₃-D, pGL₃-E, pGL₃-a, pGL₃-b, pGL₃-c and pGL₃-d, respectively; C2-R: Reverse primer for construction of luciferase reporter gene vectors pGL₃-A, pGL₃-B, pGL₃-C, pGL₃-D, pGL₃-E, pGL₃-a, pGL₃-b, pGL₃-c and pGL₃-d.</p

The cell morphology of 293T under fluorescence microscope (Olympus, micropublisher 3.3RTV, 100×) transfected by CRABP2 overexpression vector and the expression of CRABP2 detected by Westernblot.

<p>(A) Visible image transfected with lentivirus; (B) Green fluorescence image transfected with lentivirus; (C) CRABP2 expression by westernblot.</p

The expression of the <i>CRABP2</i> gene during differentiation was assessed by quantitative real-time PCR (qRT-PCR).

<p>The values were normalized to <i>GAPDH</i> mRNA expression level and the value of day 0 was set to 1. The error bars indicate the SD (n = 3).</p

Lagrange stress versus stretch in the plane of symmetry for loading velocity of 10 mms-1 (strain rate 2s-1), 100 mms-1 (strain rate 20s-1), and 500 mms-1 (strain rate100s-1).

<p>Lagrange stress versus stretch in the plane of symmetry for loading velocity of 10 mms-1 (strain rate 2s-1), 100 mms-1 (strain rate 20s-1), and 500 mms-1 (strain rate100s-1).</p

The cell morphology of C2C12 under fluorescence microscope (Olympus, micropublisher 3.3RTV, 200×) transfected by different lentivirus vector.

<p>(CON: Blank control; NC: Negative control; OE: Overexpression).</p

Transcriptional activation of the <i>CRABP</i>2 promoter was effected by <i>MyoD</i> and <i>Sp</i>1.

<p>(A) Transcriptional activation of the <i>CRABP2</i> promoter was potentiated by the MyoD expression plasmid. The empty vector or the core promoter plasmid pGL₃-E was cotransfected with the MyoD expression plasmid pcDNA3.1-MyoD into C2C12 cells. The over-expression of MyoD increased the <i>CRABP2</i> promoter activity (n = 3); (B) The transcriptional activation of the <i>CRABP2</i> promoter was repressed by the Sp1 site-directed mutation vector. The Sp1 binding site was required for <i>CRABP2</i> promoter function, and the transcription factor Sp1 facilitated the transcriptional activation of the <i>CRABP2</i> gene. Values represent the mean ± SD of three independent experiments.</p

There is a significant difference between maximum Lagrange stress (Mean±SD) of each two loading velocity.

<p>There is a significant difference between maximum Lagrange stress (Mean±SD) of each two loading velocity.</p