Overrepresented Gene Ontology terms for the differentially expressed genes in the hind legs of worker and queen pre-pupae.

<p><b>A:</b> Classification according to Biological Processes at GO. <b>B:</b> Classification according to Molecular Function. Blue: queens; Orange: workers.</p

Chart of immunolocalization of Ubx in hind legs of pre-pupae and white-eyed pupae of honeybee castes.

<p><b>A:</b> pre-pupae workers’ hind leg; <b>B:</b> white-eyed pupae workers’ hind leg; <b>C:</b> pre-pupae queens’ hind leg; <b>D:</b> white-eyed pupae queens’ hind leg. In blue: DAPI; in red: Ubx, note that the different red degrees represent the Ubx level observed in the stainings; Tar: tarsi; Btar: basitarsi; Tb: tibia; Fm: femur; Cx-Tr: coxa and trochanter.</p

Immunolocalization of Ubx (FP6.87 antibody) in honeybee prepupal hind legs.

<p><b>A-F:</b> Ubx is expressed in the tibia and basitarsi of worker pre-pupae. Note that nuclei that do not express Ubx are arranged in a similar pattern to that of bristles in the adult hind leg (arrowhead) (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040111#pone-0040111-g001" target="_blank">Figure 1D</a>). <b>G-I:</b> Ubx is expressed only in the basitarsi of queen pre-pupae hind legs. In blue: DAPI; in red: Ubx; Tar: tarsi; Btar: basitarsi; Tb: tibia. Original scale bars of confocal system.</p

Characteristics of stages of female honeybee larval development.

<p>W: workers;</p><p>Q: queens.</p

Immunolocalization of Ubx (FP6.87 antibody) in honeybee white-eyed pupale hind legs.

<p><b>A-C:</b> Ubx is expressed in the tibia and basitarsi of workers. There is a region in the tibia (which may be the future corbicula) that does not express Ubx. <b>D-I:</b> In the basitarsus and distal portion of the tibia (arrowhead) in workers, there are double nuclei that do not express Ubx, arranged in a similar pattern as that of the bristles in the adult hind leg. <b>J-L:</b> In the hind legs of queen white-eyed pupae, Ubx is expressed only in the basitarsi. In blue: DAPI; in red: Ubx; Btar: basitarsi; Tb: tibia. Original scale bars of confocal system.</p

Transcriptional pattern of <i>abdominal-A</i>, <i>ataxin-2</i>, <i>cryptocephal</i>, <i>dachshund</i>, <i>distal-less</i>, <i>grunge</i>, <i>Retinoic and fat acid Binding protein</i> and <i>ultrabithorax</i> during leg development in <i>A. mellifera</i> castes.

<p>Ordinates represent relative transcript levels assessed by qRT-PCR. Data were normalized by <i>ribosomal protein-49</i>. Three biological samples were analyzed in technical duplicates. L4, L5F and L5S: larval stages; L5PP: pre-pupae; Pw: white-eyed pupae; *: significant statistical differences between castes (P<0.01).</p

Primer sequences, annealing temperatures and fragment lengths (bp).

<p>Primer sequences, annealing temperatures and fragment lengths (bp).</p

SEM images showing divergent morphologies of <i>A. mellifera</i> worker and queen hind legs during pupal development (Pb).

<p><b>A:</b> Worker hind leg external surface, note the bristle arrangement forming the pollen basket in the tibia, i.e., corbicula. <b>B:</b> Distal portion of the tibia of worker hind leg external surface. <b>C:</b> The single bristle on the worker hind leg external surface. This bristle may be a mechanoreceptor like the other bristles on the tibia of the worker hind legs. <b>D:</b> Queen hind leg external surface. <b>E:</b> Distal portion of the tibia of the queen hind leg external surface. <b>F:</b> A bristle on the queen hind leg external surface. Arrow points to bristle socket and arrowhead points to the structure of the cuticle. Original scale bars of scanning electron microscopy system.</p

Expression of JH biosynthesis pathway genes in tissues from forager bees.

<p>Genes are grouped according to the function of the corresponding enzymes: (A) basal mitochondrial metabolism; (B, C) mevalonate pathway; (D) JH-specific steps; (E) methyl moiety transfer. Total RNA extracted from <i>corpora allata</i>-<i>corpora cardiaca</i> complexes (CA-CC), brain, fat body (FB), and ovaries were used for transcript quantification by real time RT-PCR (RT-qPCR). Each column in the graphs represents transcript levels in a single sample of 20–25 pooled CA-CC complexes, and 10 pooled brains, fat bodies and ovaries. The highest expression value for each gene was converted to 1. In the insert (C) the highest expression level among <i>fpps</i> genes was converted to 1.</p