Fig. 1: Long-term LEPR+ HSCs from middle-aged mice expanded in frequency with an accelerated decline in function as compared to young mice.
From: Age-related decline in LEPR+ hematopoietic stem cell function
A, B FACS analyses of freshly isolated total bone marrow (BM) cells from young and middle-aged C57BL/6 J (n = 8–10). Percentages of LEPR+ versus LEPR- cells within SLAM-HSCLT (LSK Flt3-CD150+CD48-) and SLAM-HSCST (LSK Flt3-CD150-CD48-), respectively. C–F Equal numbers of freshly sorted LEPR+LSK vs. LEPR-LSK cells from young or middle-aged C57BL/6 J femur BM were used at different doses (shown here dose 1000) for limiting dilution assay (LDA). Donor chimerism in peripheral blood (PB) month 1, 2, 4 and BM month 4, respectively. G Poisson statistical analysis from the LDA. Solid lines represent the best-fit linear model for each data set; dotted lines represent 95% confidence intervals. Symbols represent the percentages of negative mice for each cell dose. This plot has been modified from the original for clarity purpose. H No. of CRUs per one million transplanted cells calculated from (G); line representing median; box representing 95% confidence interval. I, J For secondary transplant, two millions of total BM cells pooled from primary hosts were intravenously injected into lethally irradiated secondary hosts. Donor chimerism in PB and BM at month 4, respectively. K, L Equal numbers of freshly sorted LEPR+ SLAM-HSCs (LSK CD150+CD48-) vs. LEPR- SLAM HSCs from young or middle-aged C57BL/6 J femur BM were used at different doses for an LDA. K Poisson statistical analysis from the LDA. Solid lines represent the best-fit linear model for each data set; dotted lines represent 95% confidence intervals. Symbols represent the percentages of negative mice for each cell dose. This plot has been modified from the original for clarify purpose. L No. of CRUs per one million transplanted cells calculated from (K); line representing median; box representing 95% confidence interval. All data are mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 using Ordinary Two-way ANOVA followed with post hoc Tukey’s multiple comparison test. N = 2 independently repeated experiments.
