Fig. 8: Heart tissue-derived CD34+ cells from ApoE-/- mice differentiated into fibroblasts.
a, b Heart-derived CD34+ cells of ApoE-/- mice treated with different concentrations of AngII (0 μM, 10 μM, 20 μM, and 50 μM) at different times (day 0, day 1, day 3, and day 5). a mRNA levels of fibroblastic markers; GAPDH was used as an internal control. Data represent mean ± SEM, n = 6. P < 0.05 was considered to be statistically significant. b Protein expression of fibroblastic markers after treatment with AngII, data represent mean ± SEM, n = 4. P < 0.05 was considered to be statistically significant. c Immunofluorescence staining of POSTN and Vimentin in tdTomato positive cells after 3 days of AngII stimulation (50 μM) (Scale bar= 50 μm). d Heatmap of the significantly changed genes (P < 0.01) discovered by the BEAM function from monocle in the branch point 3 in Fig. 4a. Cd34, ly6a, and Pi16 genes were detected in Gene Module 1, and the FABP4 gene in module 3. e–g CD34+ cells were transfected with FABP4 siRNA and then treated with AngII (50 μM) for 3 days. Data represent mean ± SEM. e Protein expression of FABP4 was determined by western blotting; f Total triglyceride in the cells, n = 3. g Protein expression of fibroblastic markers was determined by western blotting; h–k CD34+ cells were treated with PPARγ agonist pioglitazone (10 nM) and PPARγ antagonist GW9662 and then treated with AngII (50 μM) for 3 days: h mRNA levels of fibroblastic markers and ECM proteins; GAPDH was used as an internal control, n = 6; P < 0.05 was considered to be statistically significant. i Protein expression of Akt and GSK3beta was determined by western blotting. j Protein expression of FABP4 and fibroblastic markers were determined by western blotting. k Quantitative data of FABP4 and fibroblastic markers is shown; GAPDH was used as an internal control, n = 5. P < 0.05 was considered to be statistically significant.
