Fig. 6
From: TGF-β1-triggered maladaptive bone marrow endothelium impedes hematopoietic recovery

Hyperactive TGF-β1 pathway in BM ECs from PGF patients. a Heatmap of the selected TGF-β1 pathway genes in BM ECs isolated from PGF patients versus GGF patients. b GSEA showing enrichment of the epithelial–mesenchymal transition pathway in BM ECs from PGF patients. c To detect intracellular TGF-β1 and TGF-βRI levels in precultured BM mononuclear cells, BM ECs, which typically express CD309, CD34, and CD133, were first gated by using flow cytometry. Quantification of the intracellular levels of TGF-β1 (d, N = 15 per group, n = 1 per sample) and TGF-βRI (e, N = 6 per group, n = 1 per sample) levels in gated precultured BM ECs from PGF, GGF and HD was performed via flow cytometry, and the MFI was determined. The cultured BM ECs from the PGF or GGF patients were treated with LY2157299 (LY), PTN or control solvent. f Representative images (left panel; scale bars represent 50 μm) of migrated BM ECs following the indicated treatment (original magnification, ×10) and quantification (right panel) of migrated BM ECs (N = 6 per group, n = 3 per sample). g Representative images (left panel; scale bars represent 200 μm) and quantification (right panel) of tube formation of BM ECs after the indicated treatment (N = 6 per group, n = 3 per sample) (original magnification, ×4). N represents biological replicates; n represents technical replicates. The data are presented as the means ± SEMs. Statistical analyses were performed using the Mann–Whitney U test, unpaired t test and paired t test. BMMNC bone marrow mononuclear cells, EC endothelial cell, GGF good graft function, HD healthy donor, MFI mean fluorescence intensity, PGF poor graft function, PTN pleiotrophin, SEM standard error of the mean