(A) Representative images of the indicated ESCs cultured in PD03/N2B27 for 9
days. Scale bars, 100 μm.
(B) Representative phase-contrast and fluorescent images of the indicated ESCs
cultured in N2B27 medium only (the leftmost two columns) or N2B27 medium
supplemented with 3 μM 3MB-PP1 (the rightmost two columns). ESCs were
plated at clonal density and cultured in LIF/serum for 1 day. The medium was
then changed to either N2B27 or 3MB-PP1/N2B27 and cultured for another 6 days
before the images were taken. Scale bar, 100 μm.
(C) Quantification of Sox1-GFP positive colonies from
experiments described in (B). Data represent means ± SD of three
biological replicates.
(D) Representative phase-contrast and immunofluorescence staining images of
Gsk3α−/− +
Gsk3α-L195G 46C ESCs cultured in N2B27 medium with
or without 3 μM 3MB-PP1 for 11 days. Scale bar, 100 μm.