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. 2017 Mar 27;13(3):e1006682. doi: 10.1371/journal.pgen.1006682

Fig 3. Activation of canonical Wnt pathway in mESCs reduces cell proliferation and increases the number of cells in G1 phase.

Fig 3

(A) Representative images and quantification of cell number for mESCs treated with purified Wnt3a for 2 and 3 days at the indicated concentrations (n = 3). Scale bar: 400μm. (B) Representative images and cell number quantification for mESCs treated with BIO (μM) at indicated concentrations and with DMSO (μl/ml), as control, for 1, 2 and 3 days (n = 3; BIO-treated compared to respective DMSO-treated mESCs). Non-treated (serum) cells are included. Scale bar: 400μm. (C) Quantitative representation of number of BrdU positive (BrdU+) mESCs treated with indicated BIO concentrations or DMSO for 72h, as control (n = 3). (D) Cell cycle FACS analysis after propidium iodide staining of mESCs treated with BIO or DMSO for 72h (n = 3; BIO-treated compared to respective DMSO-treated mESCs). (E) Cycling index (S+G2M/G0G1) of experiments shown in D (n = 3). (F) mESCs were modified to express the FUCCI G1 phase reporter. Cells were treated for 72h either with PBS, 300ng/ml or 600ng/ml purified Wnt3a or alternatively with DMSO (0.06%) or 3 and 6 μM of CHIR99021 and analysed by FACS (Three technical replicates). All pooled data are represented as means ± SD. The asterisks indicate statistical significance by two-tailed Student’s t-test analysis (* p<0.05; ** p<0.01; ***p<0.001).