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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 ?m. (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 ?m. 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 ?m. (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.