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Fig. 4 | Journal of Hematology & Oncology

Fig. 4

From: Treatment-induced arteriolar revascularization and miR-126 enhancement in bone marrow niche protect leukemic stem cells in AML

Fig. 4

TNFα induced vascular remodeling of the leukemic BM niche via EC miR-126 downregulation. a miR-126 levels in BM ECs from normal wt and MllPTD/WT/Flt3ITD/ITD AML mice by Q-RT-PCR (n = 5 mice per group). b miR-126 levels in BM ECs from wt recipient mice engrafted with BM cells from normal wt or MllPTD/WT/Flt3ITD/ITD AML mice by Q-RT-PCR (n = 5 mice per group). c Levels of pri/pre- and mature miR-126 by Q-RT-PCR (n = 5 mice per group; left) and mature miR-126 expression by miRNA staining (right; one of three independent experiments with similar results is shown) in BM ECs from normal wt mice exposed in vitro to mrTNFα (1 ng/ml) or vehicle for 24 h. Scale bars represent a size of 10 µm. d Levels of Vcam1 and Spred1 in BM ECs from normal wt mice exposed in vitro to mrTNFα (1 ng/ml) or vehicle for 24 h by Q-RT-PCR (n = 5 mice per group). e Levels of pri/pre- (left) and mature miR-126 (right), as analyzed by Q-RT-PCR, in BM ECs from normal wt mice treated in vivo with mrTNFα (1 µg/day, ip, 3 weeks). f CD31 (FITC) and Sca-1 (PE) IF staining of tibias (left) and flow cytometry analysis of BM EC Sca-1high and Sca-1low subfractions (right) from normal wt mice treated in vivo with vehicle, mrTNFα (1 µg/day, ip), or mrTNFα+ miR-126 mimics (20 mg/kg/day, iv, 3 weeks; n = 3 mice per group). Yellow arrows indicate CD31+Sca-1high EC lined-vessels; white arrows indicate CD31+Sca-1low EC-lined vessels. Scale bars represent a size of 100 µm. One of three independent experiments with similar results is shown. g CD31 (FITC) and Sca-1 (PE) IF staining of tibias from normal wt recipient mice engrafted with BM MNCs from congenic normal mice (normal to wt) or leukemic mice (AML to wt) and of tibias from normal Spred1ECΔ/Δ recipient mice engrafted with BM MNCs from congenic leukemic mice (AML to Spred1ECΔ/Δ). Yellow arrows indicate CD31+Sca-1high EC lined-vessels; white arrows indicate CD31+Sca-1low EC-lined vessels. One of three independent experiments with similar results is shown. Results represent mean ± SEM. Significance values: *p < 0.05; **p < 0.01; ****p < 0.0001

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