<i>tan</i> is involved in female abdominal pigmentation plasticity.

<p>(A) Cuticles of control (<i>w</i>^<i>1118</i>) and <i>tan</i> mutant females (<i>t</i>^{<i>d07784</i>}) grown at 18°C, 25°C and 29°C. (B) Reaction norms of the same genotypes (n = 10 per condition). The pigmentation value corresponds to the first component of a principal component analysis of pigmentation in segments A5, A6 and A7 that captures more than 95% of the total variance. There is a significant decrease in thermal plasticity of abdominal pigmentation in <i>tan</i> mutant females. Statistical test: two-way ANOVA. ***: p<0.001.</p

<i>trx</i> is involved in female abdominal pigmentation, whereas <i>trr</i> and <i>Set1</i> are not.

<p>(A, B) When using the <i>y-Gal4</i> driver and <i>UAS-RNAi</i> transgenes at 25°C, <i>trx</i> down-regulation induces abdominal depigmentation, whereas <i>trr</i> or <i>Set1</i> down-regulation does not. In A, the effect of the RNAi transgenes against <i>trr</i>, <i>Set1</i> or <i>trx</i> (<i>VALIUM RNAi</i> lines) was compared to that of an RNAi transgene against <i>GFP</i> inserted at the same site in the same genetic background. In B, the RNAi line against <i>Set1</i> (VDRC line) driven by <i>y-Gal4</i> was compared with females heterozygous for the transgene. (C) <i>trx</i> down-regulation during late pupal stage (<i>pnr-Gal4</i> driver in combination with <i>tub-Gal80</i>^<i>ts</i> transgene) induced abdominal depigmentation. Dashed lines mark left borders of the <i>pnr</i> driver expression domain. The <i>UAS-RNAi-GFP</i> transgene is used as a negative control.</p

Temperature regulates the activity of an abdominal epidermis enhancer of <i>tan</i>, <i>t_MSE</i>.

<p>(A) <i>tan</i> genomic region (after Flybase, <a href="http://flybase.org/" target="_blank">http://flybase.org/</a>) showing the location of <i>t_MSE</i> between the genes <i>CG15370</i> and <i>Gr8a</i>. (B, C) The activity of <i>t_MSE</i> (<i>t_MSE-nEGFP</i> reporter transgene) in abdominal epidermes of young adult females is modulated by temperature. (B) nEGFP fluorescence in abdominal epidermes at 18°C and 29°C. Fluorescence on the left part of the tissue corresponds to the pleura. (C) Quantification of nEGFP fluorescence in A5, A6 and A7 hemi-tergites at 18° and 29°C (n = 10 <i>per</i> temperature). nEGFP intensity is higher at 18°C than at 29°C (t-test; ***: p<0.001).</p

Modulation of <i>tan</i> expression is necessary and sufficient for female abdominal pigmentation plasticity.

<p>Genetic manipulation of <i>tan</i> with the <i>pnr-Gal4</i> (A) or the <i>y-Gal4</i> (B) driver shows that modulation of <i>tan</i> expression plays a major role in thermal plasticity of female abdominal pigmentation. Left (A and B): <i>tan</i> down-regulation at 18°C (<i>UAS-RNAi-t</i> transgene) is sufficient to reduce pigmentation. Right (A and B): <i>tan</i> over-expression at 29°C (<i>UAS-t</i> transgene) is sufficient to increase pigmentation. In (A), dashed lines mark left borders of the <i>pnr</i> driver expression domain.</p

Temperature dramatically modulates the expression of the pigmentation gene <i>tan</i> in posterior abdominal epidermes of females.

<p>(A) Cuticle pigment synthesis pathway [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006218#pgen.1006218.ref028" target="_blank">28</a>]. Enzymes are indicated in red. (B) Quantification of pigmentation gene expression in posterior abdomen epidermes (segments A5, A6 and A7) from female <i>w</i>^<i>1118</i> pharates (left) and young <i>w</i>^<i>1118</i> adult females (right) grown at 18°C or 29°C (pools of 50 epidermes for pharates and 30 epidermes for adults, n = 3, error bars: standard deviations; gene expression at 18°C has been normalized on gene expression at 29°C). The expression of <i>tan</i>, <i>ebony</i>, <i>DDC</i>, <i>yellow</i> and <i>black</i> is moderately modulated by temperature in pharates, whereas only <i>tan</i> is dramatically modulated in young adults (t-test: *: p<0.05; **: p<0.01). The expression of <i>Tyrosine Hydroxylase</i> (<i>TH</i>) and <i>Laccase 2</i> is modulated neither in pharates nor in adults. (C) Analysis of <i>tan</i> expression in abdominal epidermes from young <i>w</i>^<i>1118</i> adult females grown at 18°C or 29°C. Note that <i>tan</i> is more strongly expressed in the posterior abdominal epidermis at 18°C than at 29°C. (D) Adult cuticle (left) and <i>tan</i> expression in abdominal epidermis (right) from females in which <i>tan</i> was down-regulated using the <i>pnr-Gal4</i> driver and a <i>UAS-RNAi-t</i> transgene. The dashed line marks the limit between the <i>pnr</i> driver expression domain (a dorsal strip) and the lateral region used as an internal control. Note the loss of pigmentation (left panel) and the strong decrease in <i>tan</i> expression (right panel) in the dorsal region, showing specificity of <i>tan</i> antisense probe.</p

Decreasing <i>EloC</i> expression suppresses ectopic veins induced by <i>blistered</i> loss-of-function.

<p>The upper allele was brought by the mother. The number of <i>EloC</i>/<i>bs^EY23316</i> females with ectopic veins was compared to the number of <i>+/bs^EY23316</i> females with ectopic veins (z-test, ^a p<0.001). The mild ectopic vein phenotype corresponds to presence of ectopic veins distal to the posterior cross-vein (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0077592#pone-0077592-g005" target="_blank">Figure 5H</a>), whereas the strong ectopic vein phenotype corresponds to presence of ectopic veins everywhere in the wing (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0077592#pone-0077592-g005" target="_blank">Figure 5G</a>).</p

Can pre-implantation biopsies predict renal allograft function in paediatric renal transplant recipients ?

<p>The upper allele was brought by the mother. The number of females with ectopic veins among flies transheterozygous for <i>Elo</i> and <i>corto</i> mutations was compared to the number of females with ectopic veins among flies with a <i>corto</i> mutation only (z-test, ^a p<0.001).</p

<i>Elo</i> genes control wing cell identity.

<p>(A): Wing from control <i>w¹¹¹⁸</i> fly (L1-L5: longitudinal veins; ACV and PCV: anterior and posterior cross-veins). (B, C): Wings from <i>+/EloB^EP3132</i> and <i>EloB^EP3132/Df(3R)BSC518</i> flies exhibit truncated L5. (D): Wings from <i>+/sd::Gal4</i> flies have a very faint ectopic vein phenotype and no margin phenotype. (E, F): Wings from flies over-expressing <i>EloA</i> exhibit ectopic vein and margin phenotypes. (G, H, I): <i>EloC^SH1520</i> and <i>EloC^SH1299</i> loss-of-function alleles diminish expressivity of the ectopic vein phenotype induced by the <i>bs^EY23316</i> loss-of-function allele. Strong phenotype: ectopic veins everywhere in the wing (shown in G). Mild phenotype: ectopic veins under the posterior cross-vein only (shown in H).</p

Down-regulation of <i>EloC</i> by RNA interference impairs both cell proliferation and cell differentiation in wing imaginal discs.

<p>(A): Clones expressing the <i>ValEloC</i> transgene (GFP⁺ cells, shown by white arrows) are located at the periphery of the disc and are very small compared to control clones. (B, C, D): Wings from pharates in which <i>ValEloC</i> is driven by <i>nub::Gal4</i> (C) or <i>rn::Gal4</i> (D), both expressed in the wing pouch <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0077592#pone.0077592-StPierre1" target="_blank">[66]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0077592#pone.0077592-Ng1" target="_blank">[67]</a> are small compared to wild-type pharate wings (B) and exhibit severe wing blade defects. By contrast, longitudinal veins (shown by asterisks) are formed in the proximal-most part of the wing blade where <i>nub::Gal4</i> and <i>rn::Gal4</i> are not expressed.</p

Deregulation of <i>EloA, EloB</i> or <i>EloC</i> expression using <i>P</i>-element insertion lines.

<p>(A): Structure of <i>EloA</i>, <i>EloB</i> and <i>EloC</i> genes showing localization of the <i>P</i>-elements used in this study. Exons are represented by boxes, and introns by lines. Black arrowheads show positions of primer pairs used to quantify <i>Elo</i> gene expression. (B): Quantification of <i>Elo</i> gene expression in <i>EloA^G4930</i>, <i>EloB^EP3132</i>, <i>EloC^SH1520</i> or <i>EloC^SH1299</i> homozygous or heterozygous larvae. (C): Quantification of <i>Elo</i> gene expression in <i>da::Gal4>>EloA^G4930</i> or <i>da::Gal4>>EloB^EP3132</i> larvae. (D): Quantification of <i>EloC</i> expression in <i>da::Gal4</i>>><i>ValEloC</i> embryos. Relative <i>Elo</i> expression levels were obtained by normalization to <i>Rp49</i> (black bars, B to D), <i>RpL12</i> (grey bars, B, C) or <i>eIF-2α</i> (grey bars, D).</p