Extended Data Fig. 4: Anti-Gremlin1 antibody upregulates the leukemic activity of CML LT-LSCs in the spleen of SCLtTA/BCR-ABL mice.
a. Schematics of the experimental design. DOX was withdrawn from the drinking water of SCLtTA/BCR-ABL mice to induce CML. Control IgG or anti-Gremlin1 antibody was intraperitoneally administered to the mice 3 days later. On day 32, equal number of spleen LT-LSCs (1000) from control IgG (qst-IgG) or anti-Gremlin1 antibody (qst-Ab) treated SCLtTA/BCR-ABL mice were sorted by FACS and transplanted to lethally irradiated recipient wild-type mice. The recipient mice were sacrificed 8 weeks after transplantation to evaluate CML engraftments. b. Body weight change (normalized to day 1) of qst-IgG and qst-Ab groups (n = 7 mice per group). Statistical analysis was performed using two-sided two-way ANOVA. c. Representative image of spleen, and spleen weight to body weight ratio of qst-IgG and qst-Ab groups (n = 7 mice per group). d. BM cellularity normalized to body weight ratio of qst-IgG and qst-Ab groups (n = 7 mice per group). e. FACS profiles and quantifications of CD45.2+ (upper) and CD45.2+Mac1+ Gr1+ (lower) cell percentage in PBMCs of recipients transplanted with BM cells from qst-IgG and qst-Ab groups (n = 7 mice per group). f,g. FACS profiles and quantifications of CD45.2+Mac1+Gr1+ (f) and CD45.2+LSK (g) cells in the spleen of qst-IgG and qst-Ab groups (n = 7 mice per group). h,i. FACS profiles and quantifications of CD45.2+Mac1+Gr1+ (h) and CD45.2+LSK (i) cells in the BM of qst-IgG and qst-Ab groups (n = 7 mice per group). Two-sided unpaired Student’s t-test was used for the statistical analysis. Error bars represent mean ± s.e.m. Numbers in FACS plots represent the mean percentage of indicated population ± s.e.m.
