Figure 6.

Additional Inhibition of PRKCz Results in an Even Higher Percentage of OCT4-, SSEA1-, and STELLA-Positive Cells

(A and A′) After day 4 without LIF, heterozygous ES cells undergo differentiation in the presence of Gö6983, while null ES cells stay as distinct colonies in (A′).

(B and B′) Gö6983 stimulates an increase in OCT4+ populations in heterozygous EBs and an even larger OCT4+ population in null EBs in (B′, insets: green and red channels separately).

(C–D′) An even higher percentage of cells are OCT4+ (C and C′) and SSEA1+ (D and D′) with Gö6983 treatment (day 12, three independent experiments).

(E and F) More STELLA+ clusters containing a larger number of cells are present in drug-treated heterozygous EBs.

(G and H) Null EBs also have more STELLA+ clusters and cells. Drug-treated null EBs exhibit a dramatic increase in the number of STELLA+ cells.

(I–K) Some cells are double positive for STELLA and VASA in drug-treated null EBs (yellow arrows). There are also VASA-only (green arrows) and STELLA-only cells (red arrows) (three independent experiments).

(L–P) Treatment with ZIP results in an increase in OCT4+ and STELLA+ cells. ZIP treatment also results in more cells that are VASA+ (three independent experiments); n = 11 for Prkci^+/−, and n = 13 for Prkci^+/− + ZIP; n = 14 for Prkci^−/−, and n = 20 for Prkci^−/− + ZIP; eight EBs assayed for both STELLA and VASA expression).

Scale bars, 100 μm in (A and A′); 50 μm in (B and B′); and 25 μm in (E, I, and L).

See also Figure S6.