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Fig. 3
β-Catenin dependant canonical Wnt signaling is compromised in Fto deficient cells.
(A) Cytoplasmic, membrane and nuclear fractions of control (Fto+/+) and Fto knockout (Fto/) MEFs treated with control () or Wnt3a -conditioned medium (+) for 4 hours were analysed by Western blot using β-Catenin antibody. Hsp90 and c-Jun were used as loading controls for cytoplasmic and nuclear fractions, respectively. (B) Fto mRNA expression in control and Fto knockout MEFs as determined by RT Real Time PCR. The data shown represent the mean ±SEM, (n = 3) (C) Quantification of cytoplasmic, membrane and nuclear β-catenin by ELISA in control and Fto knockout MEFs treated with control or Wnt3a -conditioned medium for 4 hours. The data shown represent the mean ±SEM, (n = 5). One-way ANOVA with Tukey’s multiple comparison test was performed, ***P<0.05, NS: not significant. (D) Immunofluorescence of control and Fto knockout MEFs treated with control and Wnt3a conditioned medium for 4 hours. Scale bar indicates 20 μm. This image is representative of three separate experiments.