Extended Data Fig. 7: Loss of Vcam1 inhibits the establishment and progression of haplotype-mismatched and syngeneic MLL-AF9-induced AML.

(a) Outline of experimental strategy. (b) Representative FACS plots of BM leukaemic stem cells (LSCs) from control AML-Vcam1^fl/fl (top) and AML-Vcam1^Csf1r-iCre (bottom) primary recipients, 55 days after transplantation. (c) Histogram showing the presence of leukaemic VCAM1⁺ LSCs derived from Vcam1^Csf1r-iCre mice in the BM of moribund secondary recipient mice, 103 days post-transplant. (d-e) Analysis of blood (left graph) and BM aspirates (right graph) from primary recipients transplanted with syngeneic Vcam1^fl/fl or Vcam1^Csf1r-iCre-transduced MLL-AF9-GFP LSKs, after 115 and 130 days. The red circle shows a leukemic mouse in which AML was derived from an escaped VCAM1⁺ clone. Left panel: P = 0.0153 (P = 0.003 if the VCAM1⁺ highlighted mouse is excluded); right panel: P = 0.1011 (P = 0.03 if the VCAM1⁺ highlighted mouse is excluded) (Vcam1^fl/fl n = 10, Vcam1^Csf1r-iCre n = 8 biological replicates) (e) Histogram showing the presence of leukaemic VCAM1⁺ LSCs in the BM of the sick primary recipient mouse of syngeneic Vcam1^Csf1r-iCre AML, highlighted with red circle in (d). (f) MLL-AF9-GFP⁺ cells were incubated in the presence of anti-VCAM1 blocking antibody, isotype control or camptothecin-positive control. After 4.5 hours incubation, apoptotic cells were identified by Annexin V staining, as determined by FACS (n = 4 biological replicates). Error bars, mean ± s.e.m. Mann-Whitney tests (d) and one-way ANOVA analyses followed by Tukey’s multiple comparison tests (f). Significant P values are indicated in the figure.

Source data