Fig. 5
From: Vitamin C facilitates direct cardiac reprogramming by inhibiting reactive oxygen species
Forced expression of GATA4, MEF2C and TBX5 converts human fetal cardiac fibroblast (hFCFs) into iCMs. A Strategy for creating a direct cardiac reprogramming system with hFCFs. hFCFs carrying a stable Doxy-inducible TetO promotor, mouse GMT reprogramming factors and a mCherry tag, can be converted into iCM upon exposure to Doxy. B Representative fluorescent images of mCherry expression in hFCFs after Doxy exposure. Upon exposure to Doxy, cells started to express mCherry while not being present in control cells, reflecting the presence of GMT factors. Scale bar 200 μm. C Representative FACS images of a-actinin staining positive hFCFs upon Doxy exposure. Upon exposure to Doxy, cardiac marker a-actinin expression is induced in hFCFs while not being present in non-exposed hFCFs. D Light microscopy images of GMT or Neg-transduced hFCFs on day 5, compared to cultured cardiomyocyte progenitor cells (CMPCs). Scale bar 500 μm. E, Magnified images from (D), Scale bar 250 μm. F, G Relative mRNA expression of GMT and cardiac genes MYH6 and ACTN2 during reprogramming in GMT or Neg-transduced hFCFs at different time points. RNA samples were collected from three independent experiments. Mean values + SEM of three independent experiments is shown (n = 3). Data were analyzed with two-way ANOVA. *p ≤ 0.05 vs GMT, **p ≤ 0.01 vs GMT, ***p ≤ 0.001 vs GMT and ****p ≤ 0.0001 vs GMT. (Doxy: Doxycycline; Neg: Empty vector)