FIG. 5.

Expression of β-catenin-target cardiomyogenic genes. qPCR analysis of RNA from static and rotary EBs (25, 40, 55 rpm) was performed at day 2, 4, 6, 8, and 10 of differentiation. Expression levels of noncanonical and canonical Wnts (A, B) were similar, but were modulated by culture condition, with 25 rpm rotary conditions exhibiting decreased levels of both Wnt11 and Wnt3a. In contrast, Wnt inhibitor Dkk-1 expression was increased within rotary EBs compared with static EBs (C), also corresponding to increased mesoderm-related gene expression within rotary conditions compared with static (D, E). In addition, early cardiomyogenic markers Mef-2c and Nkx2.5 were increased by rotary orbital culture (F, G), with 25 rpm conditions resulting in increased expression of sarcomeric muscle genes α-MHC and MLC-2v (H, I). n=3, P<0.05, ^#compared with all other conditions, *compared with static, **compared with 25 rpm, and ^††compared with 55 rpm. Color images available online at www.liebertpub.com/scd