Figure 4
From: Cardiac Pacemaker Cells Generate Cardiomyocytes from Fibroblasts in Long-Term Cultures
Acquisition of cardiac phenotypes in GCFs after co-culture with SANCs. (A) GCFs stably expressing enhanced green fluorescent protein (EGFP) started beating spontaneously after 2 weeks co-culture with SANCs. Typical bright-field (a) and fluorescence (b) images of GCFs are shown. (B) Expression of cardiac marker proteins in GCFs pre-labelled with EGFP; immunofluorescence images of cTnT (a, red) or desmin (b, red) in EGFP-labelled (a,b, green) GCFs. An image for the negative control for this experiment, i.e., in the absence of co-culture with SANCs, is presented vide infra. (C) Typical GCFs with Nkx2.5 expression, as indicated by the EGFP reporter signal in the vicinity of a SANC; pEGFP/Nkx2.5BD transfected GCFs were co-cultured with SANCs for 48 h. The arrow indicates one of SANCs that were originally placed in cultures. bars, 50 µm.
