Figure 3
From: Metabolite and thymocyte development defects in ADA-SCID mice receiving enzyme replacement therapy
Immature thymocytes from ADA-deficient mice show increased expression of apoptotic markers. (A) Western blot analysis of protein lysates of thymi from P14 untreated mice (lane 1–6) and 3-month-PEG-ADA-treated mice (lane 7–8). As positive control for cleaved caspase-3 staining, unfractionated thymocytes were stimulated in vitro with the apoptosis-inducer etoposide (line 9). Anti-GAPDH stain was used as protein loading control. Full-length blots are presented in Supplementary Fig. 11. (B) FACS analysis of MACS-enriched Linneg thymocytes from untreated mice at P14. DN4 (Linneg, CD25−) and DN2 + DN3 (Linneg, CD25+) populations are gated. (C) Intracellular staining for cleaved caspase-3 of cell populations shown in (B). Unfractionated Linneg thymocytes were stimulated in vitro with etoposide as apoptosis positive control. (D) Intracellular staining for cytochrome C of cell populations shown in (B). FACS plots in (C) and (D) are representative of two replicate experiments (n = 4, 2).
