hiPSCs (19‐9‐7 T,WiCell, Madison, Wisconsin) were transduced with the puromycin gene under the α‐myosin heavy chain (MHC) promoter to enable enrichment of cardiomyocytes postdifferentiation and a double fusion construct of reporter genes; firefly luciferase (Fluc) for BLI and enhanced green fluorescent protein (GFP) to enable in vivo tracking of cell engraftment longitudinally.36 hiPSCs were plated and differentiated for 30 days using a directed differentiation protocol.37 hiPSC‐CMs enriched with puromycin and treated with tumor necrosis factor‐α (TNF‐α; 20 ng/mL for 20 minutes) or angiotensin II (ANG II, Sigma‐Aldrich; 1 μM for 24 hours) with or without TPPU (1 μM) were fixed and stained with antimyosin light chain‐2a (MLC2a), anti‐myosin light chain‐2v (MLC2v), and anti‐pERK1/2 antibodies (Cell Signaling Technology, Danvers, Massachusetts) before flow cytometric analyses as described above.
Sirish P., Thai P.N., Lee J.H., Yang J., Zhang X., Ren L., Li N., Timofeyev V., Lee K.S., Nader C.E., Rowland D.J., Yechikov S., Ganaga S., Young N., Lieu D.K., Yamoah E.N., Hammock B.D, & Chiamvimonvat N. (2020). Suppression of inflammation and fibrosis using soluble epoxide hydrolase inhibitors enhances cardiac stem cell‐based therapy. Stem Cells Translational Medicine, 9(12), 1570-1584.
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