Relationship between β-major globin transcript and erythropoietic defect in G6pdΔ EBs. RT-PCR analysis of β-major RNA expression. (A) EBs derived from WT and G6pdΔ ES cell lines stably transfected with Pallino β-actin empty vector (WT and G6pdΔ^p0) or with Pallino β-actin G6pd expression vector (G6pdΔ^pG6pd) at days 13 and 15 of differentiation. RNA from WT ES cells (WT), 2 different cell lines of G6pdΔ^p0 (Δ1^p0, Δ2^p0) and G6pdΔ^pG6pd (Δ1^pG6pd, Δ2^pG6pd) were used in this experiment. (B) EB growth in the presence of 2.5 mM NAC or 2.5 mM GSH from day 11 of differentiation. EBs were harvested at 13 or 15 days of development. RNA was extracted from EBs derived from WT ES cells (WT) and from 2 different G6pdΔ ES cell lines (Δ1, Δ2). (C) EBs derived from WT ES cells (WT) and G6pdΔ ES cells (Δ1) were treated with 100 μM broad-range caspase inhibitor z-VAD-fmk from day 11 of differentiation. After 13 and 15 days of development, the EBs were harvested and assayed for β-major RNA expression. (D) EBs derived from WT and G6pdΔ ES cell lines stably transfected with Pallino β-actin empty vector (WT^p0 and G6pdΔ^p0) or with Pallino β-actin BclX_L expression vector (WT^pBclXL and G6pdΔ^pBclXL). RNA used in this experiment was extracted from EBs derived from WT^p0G6pdΔ^p0 (Δ1^p0), WT^pBclXL, and G6pdΔ^pBclXL (Δ1^pBclXL) ES cells at days 13 and 15 of differentiation. Amplified Hprt is shown as a positive control.