TIFAB overexpression enhances LSPC OXPHOS activity. (A) GSEA of RNA-seq data comparing vector control (Con) and TIFAB-overexpressing (TIFAB OE) LSPCs isolated from leukemic mice transplanted with either empty vector or TIFAB OE KMT2A::MLLT3 leukemia cells, indicating upregulation of OXPHOS with TIFAB overexpression. (B-C) OCR (Mito Stress test) (B) and ECAR (Glycolysis Stress test) (C) in Con and TIFAB OE LSPCs (n = 5). (D-F) Mitochondrial mass (n = 6) (D), mitochondrial membrane potential (n = 7) (E), and ROS levels (n = 7) (F) were assessed using MitoTracker, tetramethylrhodamine ethyl ester (TMRE), and CellROX, respectively, in Con and TIFAB OE LSPCs. (G) Fractional enrichment of ¹³C₆-labeled intermediate metabolites in Con and TIFAB OE LSPCs, measured by GC/MS (n = 3). ∗P < .05; ∗∗P < .01. Tests used in panels D-G, Mann-Whitney U test. (H) Western blot analysis of key components of the ETC complexes I, II, III, IV, and V, as well as HNF4A and TIFAB in Con and TIFAB OE LSPCs (n = 3). ACTIN served as a loading control. FDR, false discovery rate; NES, normalized enrichment score.