(A) RT-PCR analysis of three different CPC isolations for expression of Wnts compared to adult cardiac fibroblasts [cFb].
(B) Western blot protein analysis for Wnts binding to HIS-Tagged Sfrp2 after Ni+ bead pull down from control Growth Media [GM] or CPC Conditioned Media [CM].
(C) BrdU ELISA of CPCs treated for 16 with 1 nM Wnt6. *P<0.05; n=6.
(D) Western blot analysis for nuclear β-Catenin and p-JNK in CPCs at various time points after treatment with 1 nM Wnt6.
(E) BrdU ELISA of CPCs treated with 1nM Wnt6 in the presence of increasing amounts of Sfrp2 (0–100). *P<0.05; n=6.
(F) BrdU ELISA of shControl and shWnt6 CPCs in the presence or absence of 10nM Sfrp2 or 1nM Wnt6. *P<0.05; n=6. All data shown as mean ± SD of representative experiments. Nt: No treatment.
(G) Representative images of immunofluorescence staining (top) and quantification (bottom) for Nkx2.5 expression (Red) in shControl or shWnt6 CPCs after 14 days of continuous treatment with 10nM Sfrp2. * P<0.05; 4–5 images x n=3. All data are shown as mean ± SD of a representative experiment. Nt: No treatment.