Fig. 3.

miR combo reprograms pig and dog cardiac fibroblasts into cardiomyocyte-like cells.

(A) Pig cardiac fibroblasts from the left ventricle were transfected with miR combo or the non-targeting miR negmiR. Fourteen days post-transfection, RNA was extracted and analyzed for the expression of the indicated cardiomyocyte-specific genes by qPCR. Expression values are shown relative to the house keeping gene Gapdh. N = 3. *P < 0.05, **P < 0.01, ***P < 0.001.

(B) Pig cells were fixed fourteen days post-transfection and incubated with Actn2 binding antibodies to determine the number of cardiomyocyte-like cells. Representative images are shown (scale bar 100 μm) with quantification provided. Cardiomyocyte-like cell counts were determined from individual wells (N = 5). ***P < 0.001.

(C) Dog cardiac fibroblasts from the left ventricle were transfected with miR combo or the non-targeting miR negmiR. Fourteen days post-transfection, RNA was extracted and analyzed for the expression of the indicated cardiomyocyte-specific genes by qPCR. Expression values are shown relative to the house keeping gene Gapdh. N = 3. *P < 0.05, **P < 0.01, ***P < 0.001.

(D) Dog cells were fixed fourteen days post-transfection and incubated with Actn2 binding antibodies to determine the number of cardiomyocyte-like cells. Representative images are shown (scale bar 100 μm) with quantification provided. There were two populations of Actn2+ cells: low Actn2 expression and high Actn2 expression. Cell counts were determined for the high Actn2 expression population (N = 7). ***P < 0.001.