Figure 2.

Gata1⁺ Erythroid Cells Contribute a Significant Proportion of ROS to Total-Body ROS

(A) Gata1⁺ primitive erythroid cells contribute to total ROS. Cells from stressed gata1:DsRed zebrafish were gated initially for the total cell population, then evaluated by cumulative distribution function of CellROX green ROS probe signal. The 20^th to 80^th percentiles of ROS signal were gated, and the makeup of each percentile in terms of the gata1:DsRed fraction of all cells was determined. n = 20–25 individual animals per condition showing one of three independent experiments.

(B) Makeup of each percentile of ROS probe signal in terms of % gata1:DsRed contribution showing that the highest ROS signal originates mostly from Gata1⁺ erythroid cells. One-way ANOVA indicates p value < 0.0001 for overall effect and p < 0.001 in two-way comparisons between all groups.

(C) o-Dianisidine-stained vlad tepes zebrafish at 72 hpf showing a lack of erythrocytes. Scale bars represent 500 μm.

(D) Vlad tepes zebrafish have reduced total-body ROS. Vlad tepes and phenotypically normal clutch mates were treated with 20 μg/5 mL 1-naphthol followed by flow cytometry of total-body ROS indicated by CellRox ROS probe (n = 17–22 individual animals per condition in one of two experiments). All pro-oxidant exposure times were from 24 to 72 hpf.

All data are shown as the mean ± SD, with the p value from a Student t test, unless otherwise noted.