Journal: Scientific Reports

Article Title: Myocyte-specific enhancer factor 2c triggers transdifferentiation of adipose tissue-derived stromal cells into spontaneously beating cardiomyocyte-like cells

doi: 10.1038/s41598-020-80848-3

Figure Lengend Snippet: Gata4, Mef2c, and Tbx5 expression in the beating and non-beating groups. ( a ) Left three panels, gene expression levels in the beating and non-beating group. Right panel, gene expression levels in the beating and non-beating group relative to that of β-actin. ( b ) Representative immunofluorescence images of a cluster of round cells expressing Gata4 and Mef2c on day 14. ( c ) Phase-contrast and immunofluorescence images of beating cells expressing cardiac troponin T (cTnT) and Mef2c on day 28. The arrowhead indicates a beating cell. Scale bar, 100 µm. DAPI, 4′,6-diamidino-2-phenylindole, ** p < 0.01; **** p < 0.0001 by t -test or ANOVA and Tukey’s post hoc test. ( d ) The frequency of round cells expressing Gata4 and Mef2c (10 round cells were examined per experiment, n = 13). ( e ) The frequency of beating cells expressing Mef2c and cTnT (15 beating cells were analysed per experiment, n = 4).

Article Snippet: SVF was transduced with Mouse LentiORF Particles (pLenti-C-mGFP-P2A-Puro; Origene, Rockville, MD, USA): Mef2c (NM_025282, MR226865L4V), Gata4 (NM_008092, MR227022L4V), or Tbx5 (NM_011537, MR227369L4V) for overexpression, or LentiORF Control Particles (pLenti-C-mGFP-P2A-Puro, PS100093V, Origene) as an empty vector control.

Techniques: Expressing, Gene Expression, Immunofluorescence

Journal: Scientific Reports

Article Title: Myocyte-specific enhancer factor 2c triggers transdifferentiation of adipose tissue-derived stromal cells into spontaneously beating cardiomyocyte-like cells

doi: 10.1038/s41598-020-80848-3

Figure Lengend Snippet: Overexpression of SVF with Mef2c augments the number of round cells and their transdifferentiation into CMs. ( a – d ) Phase-contrast images of SVF on day 28 after transfection on day 5 ( a , control; b , Gata4; c , Mef2c; d , Tbx5). Arrowheads indicate round cells; lv, lentivirus vector. ( e ) mRNA level of Mef2c, determined by qRT-PCR, on day 28 in SVFs transfected with the control vector and Mef2c-lv (left), or scrambled shRNA and shMef2c-lv (right), on day 5, respectively. shRNA, short hairpin RNA; shMef2c, shRNA targeting Mef2c. ( f ) Number of round cells on day 14 after introducing SVF with none (none) or the control, Gata4, Mef2c, shMef2c, or Tbx5 lentivirus vector. ( g ) mRNA level of cardiac troponin T by qRT-PCR on day 28 in SVF transfected with the control, Gata4, Mef2c, shMef2c, or Tbx5. ( h ) Relationship between the mRNA levels of Mef2c and cardiac troponin T in the SVF transduced with Mef2c. R, Pearson’s product-moment correlation coefficient. The linear interpolation formula is displayed. Scale bar = 100 µm. * p < 0.05; ** p < 0.01; **** p < 0.0001 by ANOVA and Tukey’s post hoc test.

Article Snippet: SVF was transduced with Mouse LentiORF Particles (pLenti-C-mGFP-P2A-Puro; Origene, Rockville, MD, USA): Mef2c (NM_025282, MR226865L4V), Gata4 (NM_008092, MR227022L4V), or Tbx5 (NM_011537, MR227369L4V) for overexpression, or LentiORF Control Particles (pLenti-C-mGFP-P2A-Puro, PS100093V, Origene) as an empty vector control.

Techniques: Over Expression, Transfection, Control, Plasmid Preparation, Quantitative RT-PCR, shRNA, Transduction