Fig 2: Mtb co-localizes with MSC in human pulmonary and extra-pulmonary granulomas.a CD73 staining of biopsies from granuloma-positive intestinal tuberculosis patient, showing polarization of CD73-positive cells around the submucosal macrogranulomas [×100 (left panel, scale bar = 100 µm), ×200 (Right panel, scale bar = 50 µm)]. b CD73 and Ag85B dual staining performed on lung biopsy tissue from patients with pulmonary tuberculosis showing polarization of CD73+ cells (green arrows) toward the granulomas. Ag85B-positive organisms (black arrows) are seen inside the granulomas (×100, scale bar = 100 µm). c Representative of two independent human lung biopsies, Ag85B-positive organisms (brown color) are seen inside the histiocytes (black arrows), CD73+ cells are stained with blue chromogen (green arrows) and the cells showing both positivity for CD73+ and Ag85B+ organisms have been represented by red arrows (×400, scale bar = 20 µm). Insets below show the corresponding magnified CD73+ cells showing positivity of Ab85B staining. d Immunofluorescence staining performed on formalin-fixed paraffin-embedded (FFPE) tissue of human lung biopsies from patients with known tuberculosis show green fluorescence for Ag85B+ (green arrows), red fluorescence for CD73+ cells (red arrows) and colocalization signals are marked with white arrows. The strong colocalization area is shown in the yellow inset and magnified in the panel at the right. In the further right panel, corresponding green and red channel fluorescence is shown. Scale bar is 10 µm. e FFPE tissue processed for dual IF staining show Ag85B+ only (green arrows), CD105+ MSCs (red arrow) and cells positive for both CD105 and Ag85B (white arrow). The strong colocalization area is shown in the yellow inset and magnified in the panel at the right. In the further right panel, corresponding green and red channel fluorescence is shown. Scale bar is 10 µm. Data shown in this figure are representative of seven independent experiments.

Fig 3: Cell culture and characterization of PDLSCs and BMSCs. (A) Flow cytometric analysis showing positive expression of CD73, CD90, CD105, and STRO-1 and negative expression of CD34 and CD45 in PDLSCs and BMSCs. (B) Morphology of passage 2 PDLSCs and BMSCs. Green box, high magnification of the PDLSCs. Red box, high magnification of the BMSCs. Black bar, 50 µm. (C) Representative images of single clones of PDLSCs and BMSCs. Black bar, 100 µm. (D, E) Osteogenic or adipogenic differentiation ability of PDLSCs and BMSCs assayed by alizarin red staining or oil red staining. Black bar, 25 µm. (F, G) mRNA expression levels of Runx2, Col-I, OCN, and ALP in osteogenic groups and the mRNA expression levels of Ppar?, Lpl, Cebp, and Fabp in adipogenic groups. (H) Chondrogenic differentiation ability of PDLSCs and BMSCs assayed by Alcian Blue staining. Black bar, 25 µm. (I) mRNA expression levels of Col-2 and Acan in chondrogenic groups. The data are presented as the means ± SD; n = 5. *** P < 0.001 represents significant differences in inducible group compared with control group PDLSCs. ### P < 0.001 represents significant differences in inducible group compared with control group BMSCs.

Fig 4: Permanent endoglin overexpression modifies EC physiology in vitro but does not alter sprouting. a Quantification of EC migration through the uncoated transwell with a VEGF gradient after 48 h for EA.hy926 and MLEC cells [n(Mock) = 3, n(ENG+) = 3; n(WT) = 3, n(ENG+) = 3; p (EA.hy926) = 0.0297, p (MLEC) = 0.0642]. b Quantification of EC migration through the Matrigel®-coated transwell with a VEGF gradient after 24 h for EA.hy926 and MLEC cells [n(Mock) = 3, n(ENG+) = 3; n(WT) = 3, n(ENG+) = 3; p (EA.hy926) = 0.4453, p (MLEC) = 0.0344]. c EA.hy926 scratch closure after 14.5 h in culture. d Quantification of the distance migrated by EA.hy926 cells though the scratch after 14.5 h [n(Mock) = 3, n(ENG+) = 3; p = 0.0250]. e Ratio of EA.hy926 cell counts after 72 h vs. after 8 h in culture [n(Mock) = 3, n(ENG+) = 3; p = 0.2030]. f BrdU incorporation after 24 h in MLECs [n(WT) = 3, n(ENG+) = 3; p = 0.0277]. g BrdU incorporation after 4 h in EA.hy926 cells [n(Mock) = 3, n(ENG+) = 3; p = 0.0272]. h FITC-lectin-labeled sprout growth from aortic rings isolated from WT and ENG+ mice. i Number of sprouts grown from aortic rings 2.5 days after the seedtime [n(WT) = 42, n(ENG+) = 42;p = 0.2920]. j Quantification of the volume occupied by sprouts from aortic rings [n(WT) = 25, n(ENG+) = 20; p = 0.2875]

Fig 5: Permanent endoglin overexpression does not increase growth and vascularization in tumors but prevent vessel maturation and facilitates tumor cell metastasis. a First panel: Pecam1 immunostaining in the tumor tissue showing tumor vessels. Second panel: Hematoxylin–eosin staining of tumor tissue showing characteristic blood extravasation, blood lakes and edema. Third panel: a-SMA immunostaining in the tumor tissue showing vessel maturation. Fourth panel: human endoglin immunostaining in the tumor tissue. b Weight of tumors implanted in mice after 10 days [n(WT) = 35, n(ENG+) = 26; p = 0.3329]. c Quantification of the number of Pecam1-positive vessels in tumor tissue [n(WT) = 4, n(ENG+) = 6; p = 0.8758]. d Quantification of the area filled by erythrocytes in the tumor revealed by hematoxylin–eosin staining [n(WT) = 4, n(ENG+) = 6; p = 0.0484]. e Quantification of hemoglobin concentration in the tumor tissue [n(WT) = 30, n(ENG+) = 21; p = 0.0427]. f Quantification of the ratio of vessels in the tumor tissue that is partially or totally covered by a-SMA immunostained tissue [n(WT) = 8, n(ENG+) = 13; p < 0.0001]. g Epi-fluorescence images of mouse lung lobe metastases of LLC-GFP+ tumor cells. h Quantification of tumor metastatic foci per mouse lung lobe [n(WT) = 8, n(ENG+) = 9; p = 0.0004]. i Quantification of circulating LLC-GFP+ tumor cells in mice [n(WT) = 8, n(ENG+) = 9; p = 0.0107]