For confocal microscopy HT-1080 cells were transfected at 80% confluency with 500 ng DNA using Lipofectamin LTX (Thermo Fisher Scientific) according to the manufacturer's protocol (ratio 1:2 Plasmid:Lipofectamin LTX). HEK293 c18 cells were transfected at 100% confluency corresponding to 1.2*106 cells with 2.5 µg DNA by using Opti-MEM (Thermo Fisher Scientific) and linear polyethylenimine (PEI, Sigma-Aldrich) (ratio 1:3 Plasmid:PEI) (50 (link)) for the secretion assay.
HiPSC-derived cardiomyocytes were transfected using Lipofectamin 3000 (ratio 1:2 Plasmid:Lipofectamin 3,000) (Thermo Fisher Scientific) according to the manufacturer's protocol. Four hours prior transfection the medium was changed to RPMI 1,640 (Thermo Fisher Scientific) supplemented with B-27 (#17504044, Thermo Fisher Scientific) and 10% FBS.
As negative controls only the transfection reagents and Opti-MEM were added to the cells to estimate the effects of cell toxicity due to transfection and are further indicated as non-transfected (NT).
HiPSC-derived cardiomyocytes were transfected using Lipofectamin 3000 (ratio 1:2 Plasmid:Lipofectamin 3,000) (Thermo Fisher Scientific) according to the manufacturer's protocol. Four hours prior transfection the medium was changed to RPMI 1,640 (Thermo Fisher Scientific) supplemented with B-27 (#17504044, Thermo Fisher Scientific) and 10% FBS.
As negative controls only the transfection reagents and Opti-MEM were added to the cells to estimate the effects of cell toxicity due to transfection and are further indicated as non-transfected (NT).
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