FLOT2 influences the expression of CD44 in CML and AML cells. (A) Percentage of GFP⁺ (BCR-ABL1⁺) CD11b⁺ CD44⁺ cells of all GFP⁺ CD11b⁺ cells in the PB of mice transplanted with WT (red) or Flot2 KO (blue) BM, transduced with BCR–ABL1-expressing retrovirus (P < .0001; t test; n = 4). (B) Percentage of GFP⁺ (MLL-AF9⁺) Gr1⁺ CD44⁺ cells of all GFP⁺ Gr1⁺ cells in the PB of mice transplanted with WT (red) or Flot2 KO (blue) BM, transduced with MLL–AF9-expressing retrovirus (P = .031; t test; n = 4-5). (C-D) Mean fluorescence intensity of CD44 on oncogene positive (GFP⁺) lineage negative cells from mice transplanted with WT (red) or Flot2 KO (blue) BM, transduced with BCR–ABL1- (P = .0049; t test; n = 3-4) (C) or MLL–AF9-expressing retrovirus (P = .0465; t test; n = 4-5) (D). (E-F) Immunofluorescence staining for CD44 and FLOT2 on lineage negative BCR-ABL1⁺ (E) or MLL-AF9⁺ (F) cells from the BM of WT mice with CML (E) or AML (F) showing the colocalization of both proteins. The nuclei are counterstained with DAPI. The scale bar represents 10 µm. (G) Quantification of the immunofluorescence staining in (E-F). The percentage of white CD44 and FLOT2 colocalization signals per cell was quantified in WT (red) vs Flot2 KO (blue) BCR-ABL1⁺ vs MLL-AF9⁺ Lin⁻ cells. (H) Kaplan–Meier-style survival curve of WT recipient mice transplanted with WT (solid line) or Flot2 KO (dotted line) BM, transduced with BCR–ABL1- (red line) or BCR–ABL1–CD44-overexpressing retrovirus (blue line) (Log-rank test; n = 5-7).