Notch Inhibition Cooperates with AKT1 to Enhance Cardiac Reprogramming
αMHC-GFP MEFs (or GCaMP MEFs in C) were infected with GHMT or AGHMT, and treated with DMSO (vehicle) or DAPT.
(A) Representative immunostaining images of GFP, α-actinin, and cTnT at day 15 of reprogramming. Scale bar, 200 μm.
(B) Quantification of cells positive for α-actinin and cTnT as determined by immunostaining; n = 3 biological replicates.
(C) Quantification of Ca2+ flux-positive cells in GCaMP MEFs at day 15; n = 3 biological replicates.
(D) Percentage of beating cells, relative to the number of input cells; n = 3 biological replicates.
(E) Immunoblot against the Ca2+ handling proteins ryanodine receptor (RyR) and SERCA2 at day 15 of reprogramming. Densitometric quantification is shown as the average of every replicate, relative to GAPDH.
Data are presented as mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.