rabbit anti-pax2  (Thermo Fisher)

Journal: Cell reports

Article Title: Netrin1 patterns the dorsal spinal cord through modulation of Bmp signaling

doi: 10.1016/j.celrep.2024.114954

Figure Lengend Snippet: (A–L) Chicken spinal cords were electroporated at HH stage 14 with Gfp (A–C and G–I) or different concentrations of netrin1 (50 ng, 500 ng, 1 μg) (D–F and J–L) under the control of the CAG enhancer and incubated until HH stage 24. Thoracic transverse sections were labeled with antibodies against Sox2 (red, A, B, D, and E), p27 (blue, A, C, D, and F), Lhx2 (red, G and J), Isl (red, H and K), Lhx1/5 (blue/green G, I, J, and L) and Pax2 (red, I and L). The dotted box (G–L) indicates the magnified region in the adjacent panel(s). (M) Schematic spinal cord, showing the position of the dorsal progenitor (dP) domains and post-mitotic dorsal interneurons (dIs). (N and O) Ectopic Gfp expression had no significant effect on cell fate specification. In contrast the total area occupied by Sox2 + (progenitors) or p27 + (neurons) cells was significantly reduced at all concentrations of netrin1 tested (N). The dorsal spinal cord was more profoundly affected at lower concentrations of netrin1 than the ventral spinal cord (O). n > 20 sections from four embryos from each experimental condition (i.e., GFP, 50 ng, 500 ng, and 1 μg netrin), Student’s t test. (P) The different classes of dIs can be identified by specific combinations of transcription factors. (Q) There was no significant difference ( p > 0.58) in the number of caspase + cells between spinal cords electroporated with GFP and 50 ng of netrin1. In contrast, there was significant cell death when the higher concentrations of netrin1 were electroporated. n > 20 sections from four embryos from each condition (GFP, 50 ng, 500 ng, and 1 μg netrin). Student’s t test. (R) Ectopic Gfp expression had no significant effect on dI specification. However, all concentrations of netrin1 tested significantly reduced the number of Lhx2 + dI1s, Lhx1/5 + Pax2 − dI2s, and Isl1 + dI3s in a dose-dependent manner. In contrast, only the higher concentrations of netrin1 reduced the number of Lhx1/5 + Pax2 + dI4s. n > 20 sections from six embryos from each condition (GFP, 50 ng, 500 ng, and 1 μg netrin), one way ANOVA. Probability of similarity between control and experimental groups: * p < 0.05, ** p < 0.005 *** p < 0.0005. Scale bar: 100 μm.

Article Snippet: Rabbit anti Pax2 , Invitrogen , Cat#71-6000.

Techniques: Control, Incubation, Labeling, Expressing

Journal: Cell reports

Article Title: Netrin1 patterns the dorsal spinal cord through modulation of Bmp signaling

doi: 10.1016/j.celrep.2024.114954

Figure Lengend Snippet: (A–F) Thoracic transverse spinal cord sections from either control (A–C) or netrin1 − / − (D–F). E11.5 mouse spinal cords were labeled with antibodies against Ascl1 (A and D; red, dP3–dP5), Atohl1 (A and D; green, dP1), Ptf1a (top panel, B and E; dP4), Olig2 (bottom panel, B and E; pMN), phospho-histone H3 (C and F; red; cells in mitosis [M] phase), and Tuj1 (C and F; green; neurites). The dotted box in (A) and (C) indicates position of the magnified region in the adjacent panel(s). (G and H) There was no significant difference in the number of cells in M phase (G, p > 0.50; n = >20 sections from three control and three netrin1 − / − embryos) or that were caspase + (i.e., dying) (H, p > 0.28; n = >13 sections from three control and three netrin1 − / − embryos) between control and netrin1 − / − spinal cords. (I–L) To assess for changes in post-mitotic dIs, thoracic transverse spinal cord sections from either control (I and J) or netrin1 − / − (K and L) E11.5 mouse spinal cords were labeled with antibodies against Lhx2 (I and K; green; dI1), Foxd3 (I and K; red; dI2), Isl (J and L; red, dI3, MNs), Pax2 (J and L; green; dI4, dI6, v0), and Tlx3 (J and L; blue; dI3, dI5). The dotted box (I–L) indicates the magnified region in the adjacent panel(s). (M and N) Loss of netrin1 resulted in a 25% increase in the number of Atoh1 + dP1s and an almost 2-fold increase in the area occupied by the dP1s. Similarly, the area of the dP2 domain (region bounded by the Atoh1 + and Ascl1 + domains) was increased by 60%, and the Ascl1 + dP3-dP5 domain was increased by 25%. In contrast, there was no change in the area of the Ptf1a + dP4 domain ( n > 26 sections from four embryos). (O) This increased number of progenitors did not result in a loss of dIs. Rather there was a ~30% decrease specifically in the number of dI1, dI2, and dI3s ( n > 25 sections from five embryos) but not in the intermediate dorsal populations or the ventral motor neurons (MNs). Probability of similarity between control and experimental groups: * p < 0.05, ** p < 0.005, *** p < 0.0005; Student’s t test. Scale bar: 100 μm.

Article Snippet: Rabbit anti Pax2 , Invitrogen , Cat#71-6000.

Techniques: Control, Labeling

Journal: Cell reports

Article Title: Netrin1 patterns the dorsal spinal cord through modulation of Bmp signaling

doi: 10.1016/j.celrep.2024.114954

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit anti Pax2 , Invitrogen , Cat#71-6000.

Techniques: Recombinant, Protease Inhibitor, Labeling, Expressing, In Situ, Sequencing, Software, Western Blot, Imaging