SEARCH

SEARCH BY CITATION

FilenameFormatSizeDescription
STEM_210_sm_suppinfofigure1.tif12814KSupplementary figure 1: (A) Genotyping of wt and ko alleles by PCR showed that all ko mESC lines are homozygotes for the gene of interest. (B) Wnt 1 and LRP6 ko mESC lines showed no apparent differences in colony morphology compared to wt. (C) Western blot analyses showed no difference in expression level of Oct3/4 in Wnt 1 and LRP6 ko mESC lines compared to wt.
STEM_210_sm_suppinfofigure2.tif341KSupplementary figure 2: mRNA expression of Wnt1 (A) and its target genes Axin2 and Brachyury (B) during feeder free differentiation of mESCs. mRNA levels were normalized to their corresponding control conditions: Wnt1 expression in ventral midbrain embryonic day 11.5 (VM E11:5) in A, or in wt mESCs at day 4 of differentiation. Graphs represent summary of at least 4 experiments, * p≤0.05.
STEM_210_sm_suppinfofigure3.tif706KSupplementary figure 3: Characterization of the effects of Dkk1 on Wnt/β-catenin signaling in a dopaminergic cells (A) and mESCs (B and C). (A) Dkk1 decreased the activation of the Wnt/β-catenin pathway. SN4741 cells were treated with increasing doses (ng/ml) of Wnt3a and/or Dkk1 for 2.5 hours and analyzed for level of ABC (unphosphorated, activated β-catenin) and mobility shift of Dvl3 by western blotting. (B) Dkk1 decreased the activation of the Wnt/β-catenin pathway in mESCs treated with Wnt3a. LRP6 wt and ko mESCs were treated for 2.5 hour with Wnt3a (50 ng/ml), and Dkk1 (100ng/ml or 400 ng/ml) or vehicle, as indicated. (C) wt mESCs were analyzed for LRP5/6 phosphorylation and ABC level by western blot. Cells were treated with either 0.1% BSA in PBS for 16 hours (vehicle), 500 ng/ml Dkk1for 16 hours and 75ng/ml Wnt3a during the last 4 hours (Dkk1 pretreatment), or vehicle for 16 hours plus 75ng/ml of Wnt3a and 500 ng/ml of Dkk1 during the last 4 hours (Dkk1 co-treatment). Note the lower ABC and phosho-LRP5/6 levels in the Dkk1 co-treatment compared to Dkk1 pretreatment.
STEM_210_sm_suppinfo.doc23KSupporting Information

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.