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Fig. 6 | Stem Cell Research & Therapy

Fig. 6

From: Single cell and lineage tracing studies reveal the impact of CD34+ cells on myocardial fibrosis during heart failure

Fig. 6

Heart tissue-derived CD34 + cells differentiated into fibroblasts in vitro. A-E Heart-derived CD34 + cells treated with 50 ng/mL CTGF for 2 weeks: A mRNA levels of fibroblastic markers, GAPDH was used as internal control, n = 6; B-C protein expression and quantitative data of fibroblastic markers and ECM proteins after treated with CTGF for 2 weeks, n = 5. D immunofluorescence staining of PDGFRα and vimentin in CD34 + cell (Scale bar = 50 μm). E Masson's trichrome staining shows collagen deposition in CD34 + cells with CTGF treatment(Scale bar = 200 μm). F Heatmap of the significantly changed genes (P < 0.01) discovered by the BEAM function from monocle in the branch point 3 in Fig. 3F. Cd34 and Ly6a genes were detected in Gene Module 1, wnt pathway-related genes were detected in module 3 and TGFβ pathway related genes in module 4. G Genes expression of Wnt (WNT5A, WIF1, CTNNB1) and TGFβ receptor (TGFBR1,TGFBR2) pathway in CD34 + cells with or without CTGF (50 ng/mL) stimulation at the 2-week time point, n = 6. H Protein (β-catenin, Wnt5a) expression in CD34 + cells with or without CTGF (50 ng/mL) treatment for 3, 7, and 14 days, n = 5. I-J CD34 + cells were transfected with β-catenin siRNA (10 nM) and treated with CTGF (50 ng/mL) for 2 weeks, fibroblastic markers were determined by western blotting and quantitative data was shown, n = 4. K TdTomato positive cells were isolated from the heart of TAC 12-week mice (Scale bar = 100 μm). L-N TdTomato positive cells were stimulated with TGFβ1(5 ng/mL), mRNA levels of ACTA2, POSTN, COL1A1, COL1A2, COL3A1 and PDGFRA were presented upon 7 days of TGFβ1 treatment. proteins were determined by western blotting and quantitative data was shown, n = 3. O Immunofluorescence staining of α-SMA and PDGFRα in tdTomato positive cells upon 7 days of TGFβ1 stimulation (Scale bar = 50 μm). P Myofibroblastic marker (α-SMA), fibroblastic markers (periostin, PDGFRA), and ECM proteins (collagenI, fibronectin) expression in tdTomato positive cells, which TGFβ1 receptor activity was inhibited by SB525334 (2 μM), at the 7-day time point of TGFβ1 treatment. Q Schematic of proposed origin of fibroblast and myofibroblast in heart failure, *P < 0.05; **P < 0.01; ***P < 0.001

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