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. 2015 Oct 1;8(10):1295–1309. doi: 10.1242/dmm.020594

Fig. 5.

Fig. 5.

Overexpression of SOX2 in OTX2+ Daoy and hEN cells rescues self-renewal and migration deficits. (A) pReceiver-Lv-105 vector construct used for stable overexpession of SOX2 in hEN and Daoy SHH MB cells in the presence or absence of OTX2 overexpression. (B,C) Western blots depicting overexpression of SOX2 in Daoy SHH MB (B) and hEN cells (C) in the presence or absence of OTX2 overexpression. (D-K) SOX2 overexpression in OTX2+ cells results in a significant recovery in self-renewal for both Daoy (D-G) and hEN (H-K). (D,F) Representative images of spheres over subsequent passage following SOX2 overexpression in OTX2+ Daoy. (H,J) Representative images of spheres over subsequent passage following SOX2 overexpression in OTX2+ hENs. Scale bars: 400 µm. (L-O) SOX2 overexpression in OTX2+ cells rescues cell migration deficits in both Daoy (L,M) and hEN (N,O). Error bars: s.e.m. *P<0.05, **P<0.01, ***P<0.001. (L,N) Representative images of OTX2+ Daoy (L) and OTX2+ hEN aggregates (N) in collagen over 3 days following SOX2 overexpression. Arrows denote migration front for representative aggregates. n=3 biological replicates. Scale bars: 1000 µm.