Fig. 4: Expressions of cardiac transcription factors (TFs) and its perturbation in heart organoid (HO)s. | Nature Communications

Fig. 4: Expressions of cardiac transcription factors (TFs) and its perturbation in heart organoid (HO)s.

From: In vitro generation of functional murine heart organoids via FGF4 and extracellular matrix

Fig. 4

a Immunofluorescent staining to detect Nkx2-5 (green) and Tbx5 (red) in cardiac-crescent-like structures at day 3 of HO culture (n = 8, two experiments). Scale bar: 100 μm. b Immunofluorescent staining to detect Nkx2-5 (green) and Tbx5 (red) in HOs was cultured for more than 9 days. The representative HO (n = 13 out of n = 16) showed expression similar to that in embryonic hearts (E11.5). Scale bar: 100 μm. c, d Recapitulation of cardiac perturbation during in vitro HO formation from Tbx5 KO- and Isl1 KO-ESCs. c Immunofluorescent staining to detect Nkx2-5 (green) and Tbx5 (red) at day 3 of HO culture from derived Tbx5 KO- (n = 17, left) and Isl1 KO-ESCs (n = 16, right). In the Tbx5 KO-derived HO, delayed or abnormal morphogenesis was observed with decreased Tbx5 expression and disrupted typical location of Tbx5 and Nkx2-5. In the case of Isl1 KO-derived HOs, the expression of Tbx5 and Nkx2-5 was not clearly separated. Two experiments. Scale bar: 100 μm. d Immunofluorescent staining to detect cTnT (a CM marker, green)/αSMA (an SM marker, red) at day 6 of HO culture from derived Tbx5 KO- (n = 16, two experiments), Isl1 KO- (n = 17), and WT-ESCs (n = 9). The weak expressions of cTnT and/or αSMA compared to those of control (WT) HOs at day 6. Scale bar: 100 μm.

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