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CNE_23276_sm_SuppFig1.tif28958KSupporting Information Figure 1. Supporting Figure 1. Photomicrographs of double-labeled transverse (a-k, m-s) and horizontal (l) sections through the forebrain. The detected molecules are indicated on each photomicrograph. a and b, Pax6 labeling of granule cells in the olfactory bulbs, showing that those most closely located to the glomeruli are catecholaminergic cells (TH immunoreactive) and that some of them contain CB; arrow heads point to some double-labeled cells. c-h, localization of the Pax6 cells in the telencephalic hemisphere, from rostral to caudal levels, showing their distribution relative to that of the NOS positive cells (c, f), the terminal fields formed by TH-positive fibers in the region of the nucleus accumbens and the striatum (d, e), and in relation to the Nkx2.1 cells in the ventromedial region of the bed nucleus of the stria terminalis (g) and the cholinergic cells of the basal telencephalon (h). i and j, the most rostrally located cells expressing Pax7 are scattered in the mammillary region, identified in relation to the nucleus of the paraventricular organ labeled for 5-HT around the lateral recess of the infundibular ventricle (i and the higher magnification in i'), and around TH-labeled cells (j). k, Pax6 and Pax7 cells occupy the pretectum (p1) defining the three main domains, whereas in p3 both groups of labeled cells are segregated. l, horizontal section at the level marked in k, showing the dorsorostral labeling of p3 with Pax6, and the ventrocaudal labeling for Pax7. m, n and o, the basal Pax7 cell population of p3 forms a boundary with the caudal hypothalamus, as seen with double-labeling for 5HT (m), and Nkx2.1 (n), whereas the most densely labeled Pax6 cell group in the alar p3 borders the suprachiasmatic nucleus, labeled for Nkx2.2 (o). p, the boundary between p3 and p2 is highlighted by double-labeling for Nkx2.2, and the gap free of Pax6 cells in p3 is seen filled with Nkx2.2 cells (asterisks in l and p). q, Pax7 cells in the basal p3 are located lateral to the TH cells in this segment. r, Pax6 labeling with CB highlights the boundary between the thalamus and the prethalamus and shows Pax6 cells in the epiphysis. s, the ChAT-positive cells of the dorsal habenula are traversed by fibers of the habenular commissure where Pax6 cells are scattered. The monoclonal (a, b, h, r, s) or polyclonal (c-g, l-p) anti-Pax6 antibody was used according to the requirements of the combinations. For abbreviations, see list. Scale bars = 100 μm (a, b, l, n, o, p, s), 200 μm (c-h, k, m, q, r), 50 μm (i, j). This figure corresponds to the figure 8.
CNE_23276_sm_SuppFig2.tif33080KSupporting Information Figure 2. Supporting Figure 2. Photomicrographs of double-labeled transverse sections (only d is sagittal and h is horizontal) through the brain showing the detected molecules that are indicated on each photomicrograph. a-f, in the pretectal part of p1 Pax7, Pax6 and Pax3 are distributed in the commissural and juxtacommissural domains, where they are codistributed with cells expressing GABA (b) and TH (d,f). g and h, Pax7 cells are located in the interpeduncular nucleus at the level of the superior raphe nucleus (g) and in the intermediate lobe of the hypophysis where the cells intermingle with a profuse TH fiber-labeling (h, is a horizontal section). i, Pax7 imunoreactivity in the paraphysis. j, separate labeling for Pax7 in the dorsal mesencephalon and Pax6 in the tegmentum. k-m, Pax7 in the optic tectal cells colocalizes, in part, with NOS (l) and GABA (m), arrowheads indicate some double-labeled cells. n-q, the band of Pax6 cells in the mesencephalic tegmentum is located dorsal to the cholinergic (n), catecholaminergic (o), and CB-positive (p) cells but ventral to the tegmental cells labeled for Nkx2.2 (q). r-r', Pax7 cells in the interpeduncular nucleus are mainly separated from the GABA-positive cells, although the confocal analysis reveals some double-labeled cells (arrowhead in the higher magnification in r'). s, simultaneous localization of Pax7 cells in the interpeduncular nucleus above the interpeduncular neuropil in r1. t, the rostral extent of the Pax7 cells in the interpeduncular reaches the isthmic tegmentum (r0), medial to the trochlear nucleus. u and v, a large population of Pax7 cells is located dorsally in the rostral r1, at the level of the rostral raphe nucleus (u) that extends ventromedially in the caudal part of this segment into the central gray, where Pax6 cells are located (v). The monoclonal (n-p) or polyclonal (a, c, d, f, j, q, v)) anti-Pax6 antibody was used according to the requirements of the combinations. For abbreviations, see list. Scale bars = 100 μm (a-i, l, r-u), 200 μm (k, j, v), 50 μm (r'). This figure corresponds to the figure 9.
CNE_23276_sm_SuppFig3.tif18721KSupporting Information Figure 3. Suplementary Figure 3. Photomicrographs of double-labeled sagittal (a, b), transverse (c-h, j-o), and horizontal (i) sections showing the molecules indicated on each photomicrograph. a, distinct localization of Pax7 and Pax6 cells in r1 and the caudal mesencephalon. b and c, a small group of Pax3 cells is located close to the ventricle at the level of the superior reticular nucleus. d-i, the large population of Pax7 cells in r1 is rostrally medial to the isthmic nucleus (d) and continues caudally in the superior reticular nucleus, medial to the laterodorsal tegmental nucleus (e, f, i) and in the central gray (e, g, h). j, Pax6 cells are seen in the lateral margin of the cerebellum and the central gray, at the level of the locus coeruleus. k, the Pax7 cells in the central gray are distinct from the GABA cells located in this region. l, double-labeled cells for Pax6 and calbindin are abundant in the ventral zone of the rostral octavolateral area and in the central gray (yellow cells). m, Pax7 nuclear staining does not correspond to motor neurons of the facial nucleus. n, in the rostral spinal cord, scattered Pax6 cells are located in the ventral horn, and Pax7 cells are grouped medially in the dorsal horn. o, The cell nuclei labeled for Pax6 in the ventral horn do not correspond to somatomotor neurons. The monoclonal (j, l, m, o) or polyclonal (a, b, g, n) anti-Pax6 antibody was used according to the requirements of the combinations. For abbreviations, see list. Scale bars = 200 μm (a, e, g-j), 100 μm (b, d, f, k, l, n), 50 μm (c, m, o). This figure corresponds to the figure 10.

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