Journal: Frontiers in Bioengineering and Biotechnology

Article Title: ECM proteins regulate microRNA-mediated direct reprogramming of fibroblasts into cardiomyocytes via YAP signaling

doi: 10.3389/fbioe.2026.1749865

Figure Lengend Snippet: Fibroblast reprogramming via DE-DOPE/miRcombo lipoplexes is influenced by cardiac ECM protein coatings. (a,b) Immunofluorescence images (a) and quantification (b) for Mef2C (green) in AHCFs transfected with DE-DOPE/miRcombo lipoplexes and cultured for 7 days on NC and LN, FN, CI and BM coated plates. Scale bar = 100 µm. Percentage of Mef2C positive cells was calculated by counting positive nuclei for Mef2C on total nuclei number; (c,d) Flow cytometry plots (c) and quantification (d) for cTnT-positive AHCFs transfected with DE-DOPE/miRcombo lipoplexes, after 15 days culture on NC and LN, FN, CI and BM coated plates; cTnT positive cells are shown in red; (e) Immunofluorescence images for cTnT (green) expression in cells transfected with DE-DOPE/miRcombo lipoplexes, after 15 days culture on NC and LN, FN, CI and BM coated plates. Scale bar = 100 µm. (f–h) Gene expression analysis for TNNT2 (f) , ACTC1 (g) and CACNA1C (h) using ddPCR on cells transfected with DE-DOPE/miRcombo lipoplexes, after 15 days culture on NC and LN, FN, CI and BM coated plates. Data are expressed as fold change relative to the NC condition. Data are expressed as mean ± SEM of three independent experiments. Statistical analysis was performed by 1-way ANOVA.

Article Snippet: At selected time point, AHCFs were fixed in 4% PFA (Alfa Aesar), permeabilized 0.01% Triton X 100 (Sigma-Aldrich) for 10 min, and blocked with BSA 1% in PBS for 1 h. Cells were incubated with primary antibodies against Ki67 (UM870033, UltraMAB), YAP (ab52771, abcam), Mef2C (5030, Cell Signaling), cTnT (701620, Invitrogen) and α-SAR (A7607 Sigma-Aldrich) overnight at 4 °C.

Techniques: Immunofluorescence, Transfection, Cell Culture, Flow Cytometry, Expressing, Gene Expression