Fig. 4
From: CVD-associated SNPs with regulatory potential reveal novel non-coding disease genes
Identification and characterization of ncRNAs in hiPSC-CMs A Schematic protocol for differentiation of hiPS cells into cardiomyocytes, monoculture cardiac organoids (MCOs) and self-organized cardiac organoids (SCOs) in vitro. B Heat map shows the relative mRNA expression levels of the 40 ncRNAs in the 40-day-old hiPSC-CMs, or 40-day-old human cardiac organoids (including MCOs and SCOs) treated with either 1% O2 or 3% O2 for 2 or 3 days, respectively. Data are normalized to either normoxia hiPSC-CMs, or normoxia MCOs/SCOs. The red and green colors indicate high and low expression values, respectively. Means of n = 3 biological replicates per group. C Relative mRNA expression of RP11-98F14.11, COL3A1, MMP2, TGFb1, NPPA, NPPB and MYH7 in hiPSC-CMs transduced with either GapmeR RP11-98F14.11 (GM RP11-98F14.11) or GapmeR negative control (GM Ctrl) in both normoxia and hypoxia. Data are represented as Mean ± SEM; n = 3; *P < 0.05, **P < 0.01 vs. Normoxia GM Ctrl, %P < 0.05 vs. Hypoxia GM Ctrl. D Relative mRNA expression of RPL23AP9, COL3A1, MMP2, TGFb1, NPPA, NPPB and MYH7 in hiPSC-CMs transduced with either GapmeR RPL23AP9 (GM RPL23AP9) or GM Ctrl in both normoxia and hypoxia. Data are represented as Mean ± SEM; n = 3; *P < 0.05, **P < 0.01 vs. Normoxia GM Ctrl, %P < 0.05 vs. Hypoxia GM Ctrl. E Relative mRNA expression of IGBP1P1, COL3A1, MMP2, TGFb1, NPPA, NPPB and MYH7 in hiPSC-CMs transduced with either GapmeR IGBP1P1 (GM IGBP1P1) or GM Ctrl in both normoxia and hypoxia. Data are represented as Mean ± SEM; n = 3; *P < 0.05, **P < 0.01 vs. Normoxia GM Ctrl, %P < 0.05 vs. Hypoxia GM Ctrl. F Representative images of α-actinin (red) and DAPI (blue) in 40-day-old hiPSC-CMs after GM RP11-98F14.11, GM RPL23AP9, GM IGBP1P1, GM CTD-2383I20.1, or the GM Ctrl under normoxia or hypoxia for 2 days. Scale bar is 25 µm. G Cell size of the hiPSC-CMs was assessed by the Image J. 50 cells were analyzed for each condition. Data are represented as Mean ± SEM; n = 3; **P < 0.01 vs. Hypoxia GM Ctrl. Two-tailed unpaired t-test. hiPSC-CMs, human-induced pluripotent stem cell-derived cardiomyocytes; MCOs, monoculture cardiac organoids; SCOs: Self-organized cardiac orgnoids