Fig. 3.

Gas5 represses mESC endodermal differentiation. a Typical differentiating phenotype was observed in Gas5 knockdown (KD) mouse embryonic stem cells (mESCs). The arrows in Fig. 3a indicate the typical morphology of differentiated mESCs. b Expression of pluripotency and three lineages markers (ectoderm, mesoderm, and endoderm) in Gas5 KD mESCs. c Immunofluorescence results of Gata4 (endodermal marker) and Sox2 (pluripotency marker) in Gas5 KD mESCs. d–f Teratoma formation using Gas5 KD and control mESCs, and the size of Gas5 KD and control ESC-formed teratomas was calculated. g Histological analysis (H&E staining) of control and Gas5 KD mESC-formed teratomas. h Expression of pluripotency and three lineages markers (ectoderm, mesoderm, and endoderm) in Gas5 KD and control mESC-formed teratomas. i Western blot results of Gata4 (endodermal marker), Oct4, and Sox2 (pluripotency marker) in Gas5 KD and control mESC-formed teratomas. ***P < 0.001, t test, n = 3. Error bars represent SEM of the indicated experiment replicates. DAPI 4',6-diamidino-2-phenylindole, GFP green fluorescent protein