Fig. 4: NRP1 inhibition accelerates BM vascular and hematopoietic regeneration following chemotherapy.
From: Neuropilin 1 regulates bone marrow vascular regeneration and hematopoietic reconstitution
a qRT-PCR analysis of Sema3a and Nrp1 expression in BM ECs from mice at +24 h following 5-FU chemotherapy. For each gene, expression is normalized to vehicle treatment (n = 5/condition; Sema3a: p < 0.0001, Nrp1: p = 0.1291). b Representative images of VE-cadherin+ (magenta), CD31+ (green) BM vessels in femur sections from untreated control mice and at day +7 following 5-FU chemotherapy and treatment with anti-NRP1 or IgG. Nuclei are stained with DAPI (blue). Scale bar, 50 µm; magnified view scale bar, 20 µm. c BM cell counts at day +7 following 5-FU chemotherapy and treatment with anti-NRP1 or IgG (n = 5 mice/group, p = 0.0219). d At left, representative flow cytometric analysis of CD45−VE-cad+ BM ECs within BM lin− cells from control mice and at day +7 following 5-FU chemotherapy and anti-NRP1 or IgG treatment. At right, percentages and numbers of BM ECs at day +7 following 5-FU chemotherapy and the treatments shown. Dotted lines represent numbers in control mice (n = 5 mice/group; % BM ECs p < .0001; number BM ECs p = 0.0008). e At left, representative histograms show % activated caspase 3/7+ cells within lin−CD45−VE-cad+ BM ECs at day +7 following 5-FU. At right, % activated caspase 3/7+ BM ECs at day +7 (n = 5/group; p < .0001). Dotted line represents percentages of activated caspase 3/7+ cells within lin−CD45−VE-cad+ BM ECs in non-IRR controls. f PB WBC, NEU, and LYMPHs at day +7 following 5-FU and the treatments shown (n = 5 mice/group; p < 0.0001). g At left, flow cytometry of BM KSL cells at day +7 following 5-FU and the treatments shown. At right, % BM KSL cells (n = 5 mice/group; p = 0.011). h Colony forming unit quantification from 30,000 BM cells isolated from mice at day +7 following 5-FU and IgG or anti-NRP1 treatment (n = 4 replicates, mean values +/− SEM; GM: p < 0.0001; GEMM: p = 0.0201). Statistics show GM and GEMM comparisons. a, f, h Two-way ANOVA followed by Holm-Sidak’s multiple comparison two-sided t-tests; c, d, g Student’s unpaired two-sided t-test. *p < 0.05, ***p < 0.001, ****p < 0.0001. Source data provided as a Source Data file.
