XB-IMG-144539
Xenbase Image ID: 144539
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Figure 2.
Kdm2a Regulates Non-phosphorylated β-Catenin in the Nucleus
(A) The level of a β-catenin mutant protein that cannot be phosphorylated by GSK3/CK1 was reduced by Kdm2a transfection, but not by the mutant Kdm2a.
(B) Comparison of the degradation of β-CatδN in the absence and presence of Kdm2a transfection in HEK293T cells that were treated with CHX in a time series.
(C) Quantification of (B) in triplicate. Error bars represent SEM.
(D) Kdm2a exerted different effects on total, phosphorylated, and non-phosphorylated β-catenin in HEK293T cells without or with BIO treatment.
(E) Kdm2a blocked BIO-activated reporter activity. Error bars represent SEM of four replicates. ∗p < 0.05.
(F) The wild-type Kdm2a and mutant Kdm2a with point mutations or lacking different domains generated different effects on the stability of non-phosphorylated β-catenin in HEK293T cells.
(G) The catalytic function, but not the DNA binding ability, was responsible for the repression of β-catenin-stimulated reporter activity. Error bars represent SEM of four replicates. ∗p < 0.05. ns, not significant.
(H) KDM2A knockdown in HEK293T cells upregulated total and non-phosphorylated β-catenin but did not affect the phosphorylated β-catenin.
(I) KDM2A knockdown weakened the ubiquitylation of non-phosphorylated β-catenin.
(J) Kdm2a had a different effect on total and non-phosphorylated β-catenin extracted from different fractions of RKO cells following treatment with Wnt3a. Actin marked the extracts from the cytosol and cell membrane, whereas KDM2A was used as a marker for nuclear extract.
(K) Immunofluorescence showed that Kdm2a induced the degradation of non-phosphorylated β-catenin in the nuclei of SW480 cells. DAPI staining revealed the nuclei. GFP alone was used as a control.
In (A), (B), (D), (F), (H), and the “Input” in (I), WCLs were used for IB. Image published in: Lu L et al. (2015) Copyright © 2015. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View |