Figure 3.

Protocol to Improve Cardiac Differentiation by LPAR4 Regulation

(A) Real-time PCR and FACS analyses confirmed the expression of well-known cardiac progenitor markers when LPAR4 was continuously stimulated by treatment with 10 μM LPA by the established cardiac differentiation protocol. Real-time PCR analysis at cardiac differentiation day 3 and day 7, FACS analysis at day 7. (B) LPA stimulation during the early differentiation stage of the established cardiac differentiation protocol. LPAR4 was stimulated by 1 μM and 10 μM LPA for 3 days, and the results were compared to those of the untreated group. (C) LPAR4 was stimulated by 1 μM LPA for the first 3 days of differentiation and then inhibited by a combination of the antagonists AM966 and BrP-LPA for the next 3 days, and the results were compared with those of the untreated group. Statistical analyses were performed using one-way ANOVA (Newman-Keuls). ∗p < 0.01. All experiments were conducted at least in triplicate.