Figure 5.

Terminal Differentiation of Activin-Treated Cells or SAG-Treated Cells Generates Rhodopsin-Positive Neurons or Gad65-Positive Neurons, Respectively

(A) Scheme of long-term mESC differentiation in the presence of Wnti and Activin (Wnti/Act), with or without Cyc, or with Wnti and SAG (Wnti/SAG). Activin and SAG were applied 12 hr after the start of Wnti treatments, whereas Cyc was delivered simultaneously with Wnti.

(B) qRT-PCR of retinal marker (Rax, Crx, Vsx2) expression 18 DIV. Results are normalized to samples with maximal expression levels for each marker. Error bars were obtained from the error propagation formula (n = 3 independent experiments).

(C–F) Representative images of K/l EB5 cells differentiated in the indicated conditions at 22 DIV. Rhodopsin-positive cells are indicated by arrowheads in (E). Gad65-positive neurons (27.6% GAD65-positive cells among B-III-Tub-positive neurons) are indicated by arrowheads in (F). The boxed area in (D) is shown at higher magnification in (E). The scale bars represent 30 μm (C and D), 10 μm (E), and 15 μm (F).

See also Figure S7.