A) EB formation was initiated from H7 hES cells using Aggrewell plates as described under Materials and Methods. Representative images of H7 hES cells and EBs after 3, 6 and 12 days of culture later are shown; all images recorded at same magnification (100 ×). B) Total RNA was extracted from renewing hES cells and 3, 6, and 12-day EBs. Expression of self-renewal (Oct4, Nanog) and differentiation markers (T, mesoderm; Pax6, Sox1, Msx1, ectoderm; and Sox17, endoderm) as well as SFKs (Fyn, Hck, Lck, Lyn, c-Src, c-Yes) was determined by qPCR relative to control H7 hES cells maintained in mTeSR medium. Results are expressed as the average fold change relative to the undifferentiated hES cell population ± S.E.M. (n = 3; *p < 0.05; Pairwise Fixed Reallocation Randomization Test).