Nocodazole Treatment Does Not Affect Pluripotency of hESCs
(A) Brightfield images of H9 hESCs showing cell morphology after nocodazole release. Scale bars, 400 μm.
(B) qRT-PCR analysis for pluripotency and differentiation markers in H9 hESCs through a time course of 24 hr after nocodazole release. Error bars represent ±SEM of two independent experiments.
(C) qRT-PCR analysis for pluripotency and differentiation markers in H9 hESCs at passage 2, passage 3, and passage 16 after nocodazole treatment. Error bars represent ±SEM of triplicates in an independent experiment.
(D) Representative flow cytometry analysis for OCT4 expression in H9 hESCs through a time course of 24 hr after nocodazole release.
(E) Immunostaining analysis for the expression of pluripotency markers OCT4, NANOG, and SOX2 and differentiation markers BRACHYURY, EOMES, and SOX1 in DMSO- and nocodazole-treated H9 hESCs, two passages after nocodazole release. Scale bar, 200 μm.
See also Figure S2.