Figure 2—figure supplement 1.

(A) Diagram of experiment. Embryos expressing either Tg(mitfa:eGFP) or Tg(crestin:eGFP) were treated with BMP inhibitor from 12 to 24 HPF. Following treatment, embryos were dissociated, fixed, and stained for DNA content using DAPI and analyzed via flow cytometry. Scale bar = 200 µm. (B) Flow cytometry histograms showing the percentage of cells in S/G2/M phases in crestin:eGFP-positive or Tg(mitfa:eGFP-positive cell populations in BMPi-treated embryos compared to vehicle-treated embryos. (C) Fold change of crestin:eGFP-positive cells (top) and mitfa:eGFP-positive cells (bottom) in S/G2/M phases. n = 3 biological replicates of 80–100 stage-matched embryos pooled for each condition. (D) Diagram of experiment. Embryos expressing either Tg(mitfa:eGFP) or Tg(crestin:eGFP) were treated with BMP inhibitor starting at 12 HPF. Embryos were concurrently treated with EdU from 12 to 14 HPF, 16–18 HPF, or 20–22 HPF. Following EdU treatment, embryos were fixed and assess for EdU incorporation. (E) Example image of EdU incorporation in eGFP-positive cells in vehicle- and BMPi-treated embryos. (F) Quantification of EdU incorporation in Tg(crestin:eGFP) and Tg(mitfa:eGFP) embryos at different time points, n = 62–166 eGFP-positive cells from two independent experiments (N = 2) for each condition and time point. Error bars represent mean + /- SEM, P-values were calculated using ratio paired t-test for panel C and Fisher’s exact test for panel F, n.s., not significant.