Fig 2: hiPSC differentiation into hiPSC-CM and characterization. (A) Schematic illustration of experimental pipeline applied for the differentiation of human iPSC into cardiomyocytes. The basal medium was supplemented with insulin from day 7 onwards. (B) Reverse transcription-PCR analysis revealed that cardiac gene expression was positive 30 days after the differentiation process commenced. (C) On day 15, cells were purified and re-plated onto 0.1% (wt/vol) gelatin-coated coverslips. Immunostaining for a-actinin (green) and cTnT (red) shows sarcomere organization. (Scale bar, 100 µm). (D) Representative dot plots for cTnT populations. Left peaks represent the isotype control. Numbers in plots indicate the percentage of cTnT-positive cells after day 21 of differentiation. hiPSC, human induced pluripotent stem cell; CM, cardiomyocites; ACTB, ß-actin; ACTB, ß-actin; TNNT2, troponin T type 2; TNNI3, troponin I type 3; MYL7, Myosin light chain 7; MYL2, Myosin light chain 2; hiPSC, human induced pluripotent stem cells; CM, cardiomyocte; cTnT, cardiac troponin T.