The Pax7 lineage contributions in the cranial region.

<p>LacZ/YFP expression in E15.5 Pax7-cre/ROSA reporters is detected in multiple cranial regions that include: A,B) the brain (asterisks), skull, skin and nose; A–C) in the nasal (n) region, interparietal (i), parietal (p) and frontal (f) bones of the skull (viewed in (C) with brain removed); D) frontal bone (f), meninges (m) and connective tissue (c; arrows inset). E,F) YFP reporter (green) is detected in teeth (postnatal day 5 (P5)), and the trigeminal ganglia (arrow, F) near Sox9+ cells (red, F). DAPI nuclear stain in blue. G–I) Pax7-cre/ROSA LacZ mice express LacZ in the nasal cartilage, cells of the frontonasal mesenchyme (FNM), olfactory epithelium (OE) and whisker follicles (wf). Arrowheads in (G–I) show distinct border between dense and diffuse recombination+ cells in cartilage. Note (C) has been cropped in Photoshop for aesthetics. Sections are sagittal (D–H) and coronal (I). HB –hindbrain, NC –nasal cavity, sep -septum. Scale bars in D,F,G,I-500 um; E,H, inset -200 um.</p

Pax7-cre/reporter mice for lineage tracing neural crest derivatives.

<p>A,B,D) Equivalent Pax7 expression domains are seen in the frontonasal region (arrows) and neural tube (arrowheads) of E11.5 mouse embryos from different Pax7 transgenic lines: A,D) Pax7 LacZ (LacZ stain) and B) Pax7-cre/ROSA YFP (endogenous YFP expression). C) A littermate control, lacking YFP expression. E,F) E9.5 Pax7-cre/ROSA LacZ embryos show LacZ stain in neural crest regions, such as the frontonasal region (arrow) and cells leading to the pharyngeal arches (arrowheads). Box in (E) magnified in (F). G–I) In E11.5 Pax7-cre/ROSA YFP embryos, co-expression of Pax7 (red) and YFP reporter (green) can be detected in G) the neural tube (NT), H,I) olfactory epithelium (OE) and frontonasal mesenchyme (FNM; arrows). Inset highlights boxed region. Scale bars in G–I 100 um.</p

Spatiotemporal expression of neural crest markers including Pax7 in E7.5 to E9.5 mouse embryos.

<p>In C57BL/6 embryos, immunohistochemistry detects Pax7 (red) in A,B) the E7.5 to E8.5 lateral neural folds (NF; arrows); B) the E8.5 cephalic mesenchyme (arrowhead). Dapi (blue) is a nuclear stain. C,E) and D,F) are serial sections from the same axial level at E9.5. C) Pax7 is detected in the rostral (arrow), but not D) caudal (arrowhead), dorsal neural tube, whereas F) Pax3 (green) is more highly expressed in the caudal (arrowhead) and not E) rostral (arrow) neural tube. C,D) AP2-α+ cells (green) are seen mostly in the ectoderm; rare AP2-α+ cells are seen together with Sox9+ (blue) and E,F) Sox10+ cells (blue) in the branchial arches (BA); Msx1+ cells (red) were undetected in these sections. Note that rounded cells adjacent to the neural tube are background, which appear also in the negative controls. Scale bars in A–D −100 um.</p

Limited cardiovascular tissue derived through the Pax7 lineage.

<p>A–F) Recombination+ cells (LacZ+) are found in the heart and blood vessels of Pax7-cre/ROSA LacZ mice. A) Wholemount E9.5 embryos show faint LacZ+ cells in the heart (arrow). Inset shows lower magnification of panel. B–F) E15.5 sections show small populations of LacZ+ cells (arrows) in the B) heart, thymus (asterisk and inset), C) pulmonary valve (PV), D,E) aorta (Ao) and F) inner (arrow) through outer (arrowhead) layers of the brachiocephalic artery (BA). Boxes show magnified region in separate panel. PT –pulmonary trunk. Scale bars in B −500 um; insets, C–F −200 um.</p

The Pax7 lineage outside of the neural crest.

<p>In E15.5 Pax7-cre/reporter embryos, LacZ+ Pax7 descendants are seen in: A) the skin (arrowhead) and skeletal muscle (asterisk); A,B) restricted regions of the carpal bone primordium (arrows) and C) portions of the wholemount gut (arrow). In sections, a subset of LacZ+ cells are detected in: D–F) the eye; the presumptive eyelid muscle (double asterisk), in the lens (arrowhead) and its surrounding tissue (arrow), and neural retina (asterisk); G–I) the lining of the stomach (arrow); J) the pancreas (arrow identifies presumptive acinar cells); K,L) the lining of the gut (arrow; asterisk denotes the liver); and M,N) the skin (arrow), where a distinct border distinguishes the Pax7 lineage from non-lineage Pax7 (arrowhead). O–Q) YFP+ cells visualized with non-fluorescent VIP are detected in: O) the cochlea of the ear; P) lining of lung; and Q) liver (arrows for all). Both YFP and LacZ Pax7-cre/reporters gave the same results, as did fluorescent and non-fluorescent detection methods. Boxes show magnified region in adjacent panel. Scale bars in D,G,K,O–Q are 500 um; E,F,H–J, L–N are 100 um.</p

Identification using DNA sequencing of the bacteria isolated from scorpions, their percent identity as indicated by BLAST, clade organization, and OTU assignment.

(DOCX)</p

Phylogenetic tree of Mollicutes 16S rRNA.

The phylogenetic tree was constructed using Maximum Likelihood, including a total of 312 sequences representing Mollicutes taxa. Scorpion sequences are in bold. Red collapsed regions indicate 12 sequences reported in this study from H. arizonensis and S. mesaensis. For an expanded version of this figure see S2 Fig.</p

Phylogenetic tree showing species-specific clades for telson bacteria isolated from <i>S</i>. <i>mesaensis</i> and <i>H</i>. <i>arizonensis</i>.

A Maximum Likelihood phylogeny was constructed based on the DNA sequences of 114 16S rRNA samples isolated from S. mesaensis (n = 32) and H. arizonensis (n = 82) telsons. Scorpion-specific clades are identified by vertical bars numbered 1–12 with samples from H. arizonensis indicated in black; S. mesaensis indicated in red. We considered clades to be scorpion-specific if they contained 3 or more taxa from a single scorpion species and were supported by bootstrap values greater than 70.</p