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. Author manuscript; available in PMC: 2024 Feb 16.
Published in final edited form as: Cell. 2023 Feb 16;186(4):732–747.e16. doi: 10.1016/j.cell.2023.01.020

Figure 5. Inhibition of ferroptosis rescues HSC depletion due to MYSM1 loss.

Figure 5

(A) Quantification of CD34+CD45RA−, ST-HSC, and LT-HSC populations in AAVS1 or MYSM1 edited cells, and MYSM1 edited cells treated with indicated chemicals. (B) Quantification of mean fluorescence intensity (MFI) of cellular ROS level of sorted CD34+CD45RA−CD90+ cells with AAVS1 or MYSM1 editing, and MYSM1 edited cells with indicated chemical treatments measured by CellROX dye. (C) Quantification of cellular lipid peroxidation level of sorted CD34+CD45RA−CD90+ cells with AAVS1 or MYSM1 editing, and MYSM1 edited cells with indicated chemical treatments measured by ratio of oxidized and non-oxidized BODIPY dye. (D) Quantification of mean fluorescence intensity (MFI) of O-Propargyl-puromycin based translation rate analysis on CD34+CD45RA−CD90+ sorted cells with AAVS1 or MYSM1 editing, and MYSM1 edited cells with indicated chemical treatments. (E) Schematic of in vivo rescue xenotransplantation experiment. (F) Percentage of total human CD45+ cells after RBC depletion from peripheral blood of NBSGW mice xenotransplanted with AAVS1 and MYSM1 edited cord blood CD34+ HSPCs treated with either vehicle (3%DMSO) or 20mg/kg Liproxstatin-1. Peripheral blood was harvested through retro-orbital plexus and analyzed at 4, 8 and 12 weeks post-transplantation. (G) Percentage of edits of MYSM1 edited cells before and after xenotransplantation from peripheral blood of NBSGW mice xenotransplanted with AAVS1 and MYSM1 edited cord blood CD34+ HSPCs treated with vehicle (3%DMSO) or 20mg/kg Liproxstatin-1. (H) Percentage of total human CD45+ cells after RBC depletion from bone marrow (left) or spleen (right) of NBSGW mice xenotransplanted with AAVS1 and MYSM1 edited cord blood CD34+ HSPCs treated with either vehicle (3%DMSO) or 20mg/kg Liproxstatin-1. (I) Percentage of edits of MYSM1 edited cells before and after xenotransplantation from bone marrow (left) or spleen (right) of NBSGW mice xenotransplanted with AAVS1 and MYSM1 edited cord blood CD34+ HSPCs treated with vehicle (3%DMSO) or 20mg/kg Liproxstatin-1. (J) Cell numbers per 30,000 RBC depleted bone marrow cells of progenitor cells (CD34+), T cells (CD3+), myeloid (CD33+, CD11b+), B cells (CD19+) and granulocytes (CD66b+) of NBSGW mice xenotransplanted with AAVS1 and MYSM1 edited cord blood CD34+ HSPCs treated with either vehicle (3%DMSO) or 20mg/kg Liproxstatin-1.