Fig. 4

Biological effects of thioredoxin deletion on murine hematopoietic stem/progenitor cells. TXN1^fl/fl (control) and ROSA-CreER-TXN1.^fl/fl mice were treated with tamoxifen (75 mg/kg) i.p. daily for 5 days. Mice were sacrificed at day 10. Bone marrow cells were harvested and enriched for hematopoietic stem/progenitor cells using lineage depletion kit. The hematopoietic stem/progenitor- enriched cells were then measured for (A) beta-gal expression by flow cytometry staining, and Annexin (B). ROS measurement by carboxy-H2DCFDA staining. C JC probe to measure mitochondrial membrane polarization (D). E, F Seahorse measurement of oxygen consumption and cellular ATP concentration. G Relative mitochondrial DNA content (mtDNA:nDNA), The results were normalized to the ratio of mtDNA:nDNA of control cells. (n = 10 Mean ± SEM or SD). H–J PARK2, PINK1 and LC3B mRNA expression in murine HSPCs were quantified using qPCR (n = 3–5 Mean ± SEM or SD)