![Young vs old BM macrophages interact differently with leukemia cells. (A) Number of primary GFP+ (BCR-ABL1+) BP-1+ cells plated on macrophages from young (red) vs old (blue) mice (n = 6); 2 × 104 GFP+ (BCR-ABL1+) BP-1+ cells had been plated. The P values are as indicated (1-way ANOVA, Tukey test). (B) Number of BCR-ABL1+ BA/F3 cells plated on macrophages from mice of increasing ages (3-16 weeks) (n = 3); 2 × 104 BA/F3 cells were plated. Cell numbers were counted 5 days after plating. The P values are as indicated (1-way ANOVA, Tukey test). (C) Number of primary GFP+ (BCR-ABL1+) BP-1+ B-ALL cells from young or old mice with established B-ALL plated on young vs old macrophages (MΦ); 1 × 105 leukemia cells had been plated. The overall P value is indicated (1-way ANOVA). The data are representative of 2 independent experiments. (D) Cell cycle analysis by 4′,6-diamidino-2-phenylindole (DAPI) staining of primary GFP+ (BCR-ABL1+) BP-1+ cells plated on macrophages from young (red) vs old (blue) mice. The P values are as indicated (1-way ANOVA, Tukey test). (E) Number of primary GFP+ (BCR-ABL1+) BP-1+ cells adhering to macrophages from young (red) vs old (blue) mice (n = 3); 1 × 105 cells had been plated and were allowed to adhere for 6 hours (not significant [n.s.]; Student t test). (F) Number of primary GFP+ (BCR-ABL1+) BP-1+ cells that had migrated toward macrophages from young (red) vs old (blue) mice after 24 hours; 1 × 105 leukemia cells had been plated (P = .01; Student t test, n = 3). (G) Number of primary GFP+ (BCR-ABL1+) BP-1+ cells from 6-week-old mice plated in the CM of macrophages (MΦ) from young (red) vs old (blue) mice; 2 × 104 GFP+ (BCR-ABL1+) BP-1+ cells had been plated. The P values are as indicated (1-way ANOVA, Tukey test) (n = 6). (H) Number of primary GFP+ (BCR-ABL1+) BP-1+ cells from young (left) vs old (right) mice with established B-ALL plated in the CM of macrophages (MΦ) from young (red) vs old (blue) mice; 2 × 105 leukemia cells were plated. The overall P values are as indicated (1-way ANOVA).](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/138/19/10.1182_blood.2021011557/4/bloodbld2021011557f3.png?Expires=1724246602&Signature=vCviN1C8ccuCagd9bINY7VbDXeAnulyT6KooRG8DHI2Ga-tJpwhng3UxdhaXF2m27926rqrMcE9VlEOGSYAkQhqO3JyyGGgVwTBb4R3gUctjwQWnu5h~Vwvk3mLmDa1OD71M6MZ18DspcBbug3GC-c1YQEYC6ZxunumBu6QNiUFcBypHrFRld1~yHgrwFuXNyqWJz-y2yFIQ0IcSTk9GhSSnpBC3zpYc0VpnbFL9ouOcM17th7ooLXBlIX9P5kPRq2uHVha~O4-fpMWJQuqkIJdMLDPulvUykOXw5avZOkUaH1o9JMq~46G1fZNZJfLvcFXNA-bfEXAgDSrbGhSZ5A__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Young vs old BM macrophages interact differently with leukemia cells. (A) Number of primary GFP+ (BCR-ABL1+) BP-1+ cells plated on macrophages from young (red) vs old (blue) mice (n = 6); 2 × 104 GFP+ (BCR-ABL1+) BP-1+ cells had been plated. The P values are as indicated (1-way ANOVA, Tukey test). (B) Number of BCR-ABL1+ BA/F3 cells plated on macrophages from mice of increasing ages (3-16 weeks) (n = 3); 2 × 104 BA/F3 cells were plated. Cell numbers were counted 5 days after plating. The P values are as indicated (1-way ANOVA, Tukey test). (C) Number of primary GFP+ (BCR-ABL1+) BP-1+ B-ALL cells from young or old mice with established B-ALL plated on young vs old macrophages (MΦ); 1 × 105 leukemia cells had been plated. The overall P value is indicated (1-way ANOVA). The data are representative of 2 independent experiments. (D) Cell cycle analysis by 4′,6-diamidino-2-phenylindole (DAPI) staining of primary GFP+ (BCR-ABL1+) BP-1+ cells plated on macrophages from young (red) vs old (blue) mice. The P values are as indicated (1-way ANOVA, Tukey test). (E) Number of primary GFP+ (BCR-ABL1+) BP-1+ cells adhering to macrophages from young (red) vs old (blue) mice (n = 3); 1 × 105 cells had been plated and were allowed to adhere for 6 hours (not significant [n.s.]; Student t test). (F) Number of primary GFP+ (BCR-ABL1+) BP-1+ cells that had migrated toward macrophages from young (red) vs old (blue) mice after 24 hours; 1 × 105 leukemia cells had been plated (P = .01; Student t test, n = 3). (G) Number of primary GFP+ (BCR-ABL1+) BP-1+ cells from 6-week-old mice plated in the CM of macrophages (MΦ) from young (red) vs old (blue) mice; 2 × 104 GFP+ (BCR-ABL1+) BP-1+ cells had been plated. The P values are as indicated (1-way ANOVA, Tukey test) (n = 6). (H) Number of primary GFP+ (BCR-ABL1+) BP-1+ cells from young (left) vs old (right) mice with established B-ALL plated in the CM of macrophages (MΦ) from young (red) vs old (blue) mice; 2 × 105 leukemia cells were plated. The overall P values are as indicated (1-way ANOVA).