Locus of the <i>gsb</i> gene.

<p>(<b>A</b>) <i>gsb</i> mutant alleles. The two deficiencies, <i>Df(2R)IIX62</i> and <i>Df(2R)SB1</i>, as well as the two hypomorphic alleles, <i>gsb525</i> and <i>gsbP1155</i>, are depicted. Neighboring genes uncovered by <i>Df(2R)IIX62</i>, <i>zip</i>, <i>uzip</i>, <i>CG3441</i>, and <i>gsbn</i> upstream of <i>gsb</i>, <i>gol</i> and <i>dTKR</i> downstream of <i>gsb</i>, and their direction of transcription are indicated (the rigth telomere of the second chromosome is to the right). Exons are marked by black boxes in the enlarged portion of (<b>A</b>) and also in (<b>B</b>). (<b>B</b>) Map of <i>gsb0-</i>525 abd <i>gsb0</i>-ΔHC transgenes. Both transgenes contain the upstream epidermis enhancers of <i>gsb</i>, GEE and GLE (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030980#pone-0030980-g001" target="_blank"><b>Fig. 1A</b></a>; Li et al., 1993), the <i>gsb</i> promoter, and the entire 3′ UTR of <i>gsb</i>. In <i>gsb0</i>-ΔHC, 519 bp of coding region between the <i>gsb525</i> mutation and a <i>SacII</i> site are deleted, resulting in a shift of the open reading frame after the <i>gsb525</i> nonsense mutation. (<b>C</b>) Sequence surrounding the <i>gsbP1155</i> insertion site. The negative numbers refer to nucleotides upstream of the transcription start site. The eight nucleotides, duplicated during insertion of the P-element, are underlined.</p

Rescue of fertility of <i>gsb</i> mutant males by <i>gsb</i>-Prd and <i>gsb</i>-Pax3 transgenes.

<p>Percentage of fertile males among <i>gsb</i> mutant males that were rescued by one or two copies of <i>gsb-res</i>, <i>gsb</i>-Prd or <i>gsb</i>-Pax3 transgenes (actual numbers of fertile males per total number of rescued males are given in parentheses). Rescued males were obtained from crosses described in legend of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030980#pone-0030980-t001" target="_blank">table 1</a> and were placed individually with at least three wild-type virgin females in fresh vials to score fertility. nd, not determined.</p

Map of <i>gsb-res</i>, <i>gsb-</i>Prd and <i>gsb-</i>Pax3 transgenes.

<p>The <i>gsb-res</i> transgene corresponds to the enlarged 20-kb genomic fragment in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030980#pone-0030980-g001" target="_blank"><b>Fig. 1A</b></a>, which includes the <i>gsb</i> transcribed region as well as adjacent 14-kb upstream and 3-kb downstream sequences <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030980#pone.0030980-Gutjahr1" target="_blank">[9]</a>. The upstream sequence also contains the 5′ portion of the <i>gsbn</i> up to part of the third exon. In <i>gsb</i>-Prd and <i>gsb</i>-Pax3 transgenes, the <i>gsb</i> coding region (except of a small region encoding the C-terminus) is replaced by <i>prd</i> and <i>Pax3</i> cDNAs, while upstream and downstream regions are retained. The <i>gsb</i> intron is also retained by inserting it between sequences of the <i>gsb</i> and <i>prd</i> or <i>Pax3</i> leaders. Coding regions are indicated as black boxes except for the paired-domain (PD) and the <i>prd</i>-type homeodomain (HD) which are hatched. The <i>gsb</i> and <i>gsbn</i> introns are indicated as open boxes. The transcription start of <i>gsb</i> is marked by 0, and poly(A) addition signals AATAAA are indicated.</p

Expression of Gsb and Prd proteins and <i>Pax3</i> mRNA under control of the <i>gsb cis</i>-regulatory region.

<p>Expression of Gsb protein in wild-type embryos (<i>ry⁵⁰⁶</i>; <b>A</b>), of Prd protein in transgenic <i>gsb</i>-Prd embryos (<b>B</b>), and of <i>Pax3</i> mRNA in transgenic <i>gsb</i>-Pax3 embryos (<b>C</b>) at the extended germ band stage. Wild-type embryos were stained with anti-Gsb antiserum and transgenic embryos, collected from homozygous <i>gsb</i>-Prd or <i>gsb</i>-Pax3 stocks, were stained with anti-Prd antiserum or hybridized <i>in situ</i> with digoxigenin-labeled <i>Pax3</i> cDNA. Unfolded embryos are shown and oriented with their anterior to the left. Scale bar: 100 um.</p

Rescue of the CNS phenotype of <i>gsb</i> mutant embryos by <i>gsb</i>-Prd and <i>gsb</i>-Pax3 transgenes.

<p>Patterns of longitudinal and commissural axons in the CNS of wild-type (<i>ry⁵⁰⁶</i>; <b>A</b>) and <i>Df(2R)IIX62/gsb525</i> embryos without (<b>B</b>) and with one copy of the <i>gsb</i>-Prd (<b>C</b>) or <i>gsb</i>-Pax3 transgene (<b>D</b>). Embryos at stage 15 were collected from crosses between <i>Df(2R)IIX62/CyO</i>, <i>hb-</i>LacZ; <i>gsb</i>-Prd/+ or <i>Df(2R)IIX62/CyO</i>, <i>hb-</i>LacZ; <i>gsb-</i>Pax3/+ males and <i>gsb525/CyO</i>, <i>hb-</i>LacZ females, and double stained with rabbit antiserum against ß-galactosidase and monoclonal antibody BP102. One quarter of the embryos did not stain for ß-galactosidase as expected. Half of these embryos have missing or reduced posterior commissures as expected for <i>gsb</i> mutants, the other half displays fully rescued commissural patterns as in wild-type embryos. Scale bar: 10 um.</p

Cuticular phenotypes of <i>gsb</i> mutants.

<p>(<b>A</b>) <i>Df(2R)IIX62/CyO</i>, (<b>B</b>) <i>Df(2R)IIX62</i>, (<b>C</b>) <i>Df(2R)IIX62/Df(2R)KrSB1</i>, (<b>D</b>) <i>Df(2R)IIX62/gsb525</i>, (<b>E</b>) <i>gsb525</i>, (<b>F</b>) <i>gsbP1155</i>, (<b>G</b>) <i>Df(2R)IIX62</i>; <i>gsb0</i>-525 (<b>H</b>) <i>Df(2R)IIX62</i>; <i>gsb0</i>-ΔHC. Note in strong <i>gsb</i> mutants (<b>B</b>, <b>C</b>), the ventral naked cuticle region of each segment is transformed into denticle belt, generating an overall denticle pattern, which is in contrast to wild-type (<b>A</b>). Scale bar: 50 um.</p

Rescue of <i>gsb</i> mutant embryos to viable adults by <i>gsb</i>-Prd and <i>gsb</i>-Pax3 transgenes.

<p>Percentage of rescued <i>gsb^-</i> flies harboring one or two copies of <i>gsb-res</i>, <i>gsb-</i>Prd or <i>gsb-Pax3</i> transgenes (actual numbers of rescued flies per total number of expected <i>gsb</i> mutants are given in parentheses). <i>Df(2R)IIX62/gsb⁵²⁵</i> flies carrying one or two copies of the transgenes were obtained as offspring from the crosses between <i>Df(2R)IIX62/SM1</i>; <i>P/P</i> (P stands for the transgenes) males and <i>gsb⁵²⁵/SM1</i> or <i>gsb⁵²⁵/SM1</i>; <i>P/P</i> females. <i>Df(2R)IIX62/gsb^P1155</i> and <i>gsb⁵²⁵/gsb^P1155</i> flies carrying one copy of the transgenes were obtained from the crosses between <i>Df(2R)IIX62/SM1</i>; <i>P/P</i> or <i>gsb⁵²⁵/SM1</i>; <i>P/P</i> males and <i>gsb^P1155/SM1</i> females. nd, not determined.</p

Process of GIS cloud-computing service for iron mine potential evaluation.

<p>Process of GIS cloud-computing service for iron mine potential evaluation.</p

Adjusted associations between health insurance benefit design (comparing having both inpatient and outpatient coverage to having only inpatient coverage) and health care utilization.

<p>Note: Covariates in all models include gender, age, education level, employment status, household size, household wealth, self-assessed health, distance from home to health facilities, and county variables.</p

Resource synchronism of the node portal.

<p>Resource synchronism of the node portal.</p