Figure 3.

Affected Hematopoiesis in iPS-derived erythroid cells of TTD patients. In vitro erythroblast differentiation performed on iPS cells derived from TTD218UT (TTD) fibroblasts, control fibroblasts (control F) and peripheral blood mononuclear cell derived erythroblasts (control E).(A) Cytospins and Giemsa-Grünwald staining of in vitro differentiated erythroblasts. Arrows indicate multinucleated cells. (B) Quantification of multinucleated cells, expressed as a percentage of total cells [>500 cells, counted by two independent researchers (N = 2)]. (C) Dot plots of flow-cytometric analysis of differentiated erythroid control and TTD cells. Lower bar graph depicts the quantification of the gated dot-plots [Q8=FSC^low/SSC^low (normal size and intracellular structures); Q5=FSC^high/SSC^low (bigger cells); Q6=FSC^high/SSC^high (bigger cells, more intracellular structures); Q7=FSC^low/SSC^high (normal size, more intracellular structure); N = 5 for each. Error bars represent standard deviation] (D) HPLC analysis of different hemoglobin variants. Bar graphs depict the percentage of hemoglobin variants: “HbF total” = sum of HbF1 and HbF2, Hb-HbH = sum of all non-aberrant hemoglobin variants, HbA2, HbE, HbH represent the peaks identified as HbH, probably representing HbBarts (beta tetramers or gamma tetramers respectively) and “Hb Total” = sum of all the peaks. Experiments represent an N = 3 for TTD and N = 6 for control iPS-derived erythroid cells. Error bars represent standard deviation and student t-test was performed to calculate significance, * <0.05 and ***< 0.01.