XB-IMG-144541
Xenbase Image ID: 144541
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Figure 4.
Kdm2a Demethylates β-Catenin
(A) Immunoprecipitated non-phosphorylated β-catenin was methylated, which was reduced in response to Kdm2a transfection. IgG antibody was used as a negative control.
(B) Comparison of the methylation status between the phosphorylated and non-phosphorylated β-catenin, and their responses to Kdm2a.
(C) Loss of KDM2A function caused an increase in methylated β-catenin.
(D) The wild-type and the mutant Kdm2a showed different demethylaton activity on β-catenin.
(E) β-catenin with an N-terminal deletion was methylated and was then reduced in the presence of Kdm2a transfection.
(F) β-Catδ(275–360) was weakly methylated, which did not change in response to Kdm2a transfection.
(G) Detection of the methylation status of the isolated aa 272–390 region and its response to Kdm2a overexpression.
(H) The wild-type β-catenin and the mutant β-catenin with lysine mutations showed different methylation statuses, and Kdm2a generated different effects on their stability and methylation statuses.
(I) Kdm2a overexpression induced strong ubiquitylation in wild-type protein, but not in the mutant protein.
(J) A time-course experiment illustrating that the mutant β-catenin was resistant to Kdm2a.
(K) Quantification of (J) in triplicate. Error bars represent SEM.
(L) Methylation status of non-phosphorylated β-catenin in different cellular compartments of HEK293T cells in the absence and presence of BIO treatment. GAPDH and KDM2A were used as markers for cytosolic and nuclear extracts, respectively.
(M) Wnt3a treatment of HEK293T cells enhanced the methylation of non-phosphorylated β-catenin, which was reduced in response to Kdm2a plasmid transfection.
(N) Xenopus Kdm2a, human KDM2A, and Xenopus Kdm2b exhibited similar demethylation activities for β-catenin.
In (A)–(E), (G), and (H), “Lysate” indicates WCL, which was also used in (J). In (L), nuclear and cytosolic extracts were used for IB, as indicated. Immunoprecipitates were used for remaining 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 |