Fig. S2

Analysis of PH3+ and acridine orange (AO) + cells in aphidicolin treated embryos during radial sorting. PH3 immunolabeling in Tg(foxd3:gfp) (A–A″) and Tg(foxd3:gfp) embryos treated with aphidicolin between 45 and 48 hpf and analyzed at 48 hpf (B–B″). Arrow in A″ indicates a SC that is GFP and PH3 positive. A″, merge of A and A′; B″, merge of B and B′. Scale bar = 20 μm. (C) Quantification of the number of PH3+ cells within a defined region of the PLLn at 48 hpf in control (average of 1.1 ± 0.31, n = 10) and aphidicolin treated embryos between 45 and 48 hpf (average of 0, n = 7) (⁎⁎, p = 0.0067). Acridine orange staining at 72 hpf in control (D–D″) and aphidicolin treated embryos between 45 and 54 hpf (E–E″) within a defined region of the PLLn. Scale bar = 20 μm. Arrowheads designate the PLLn in D′ and E′. D″, merge of D and D'; E″, merge of E and E′. (F) Quantification of the number of AO positive cells in control (average of 0.25 ± 0.13, n = 12) and embryos treated with aphidicolin between 45 and 54 hpf (average of 0.33 ± 0.19, n = 12) within a defined region of the PLLn (ns, p = 0.67). (G) Quantification of the percentage of AO+ SCs relative to the total number of SCs within a defined region of the PLLn at 72 hpf in controls (average of 1 ± 0.53, n = 15 embryos), and 45–54 hpf treated embryos (average of 2.31 ± 1.03, n = 12 embryos). AO staining at 72 hpf in control (H–H″) and aphidicolin treated embryos between 45 and 54 hpf (I–I″) within a defined region of the spinal cord. H″, merge of H and H′; I″, merge of I and I′. Scale bar = 20 μm. (J) Quantification of the number of AO positive cells in control (average of 0.66 ± 0.31, n = 12) and aphidicolin treated embryos between 45 and 54 hpf (average of 20.17 ± 1.07, n = 12) within a defined region of the spinal cord (⁎⁎⁎⁎, p < 0.0001). TL, transmitted light.