Journal: Frontiers in Oncology

Article Title: Replication stress increases mitochondrial metabolism and mitophagy in FANCD2 deficient fetal liver hematopoietic stem cells

doi: 10.3389/fonc.2023.1108430

Figure Lengend Snippet: Mitophagy activity was decreased in Fancd2-deficient BM HSC. (A) Relative MFI (left panel) and representative flow cytometric plots (right panel) showing MMP in adult (8-12 weeks) HSCs (Fancd2 WT n=6 mice, HET n=2 mice, KO n=8 mice, left panel). (B) Relative MFI of mtROS in adult HSCs (Fancd2 WT n=5 mice, HET n=2 mice, KO n=5 mice). (C) Mitochondrial area, mitochondrial concentration and mitochondrial polarity in HSCs. HSCs were stained with anti-Tomm20 and analyzed by IFM (Fancd2 WT n=559 cells, KO n=431 cells). (D) Relative MFI (left panel) and representative flow cytometric plots (right panel) of mitophagy in BM HSCs (Fancd2 WT n=5 mice, HET n=2 mice, KO n=4 mice). (E) Relative MFI (left panel) and representative FACS plots (right panel) of LA in BM HSCs (Fancd2 WT n=6 mice, KO n=5 mice, left panel). All relative data are divided with average of controls (WT or HET) and show mean ± SD. *P<0.05, ****P<0.0001, ns, not significant.

Article Snippet: HSCs were sorted by FACS Aria II or III (BD) and 500 cells are seeded onto 3ml of mouse methylcellulose media (R&D Systems, HSC007) with APH (50-100ng/ml) or imTOR of Rapamycin (Thermo PHZ1235, diluted with DMSO for 10mM and used as 1:1000, final concentration 10μM) or iTgf-β of SD-208 (CAYMAN 16619, diluted with DMSO for 10mM and used as 1:1000, final concentration 10μM).

Techniques: Activity Assay, Concentration Assay, Staining