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. Author manuscript; available in PMC: 2022 Feb 14.
Published in final edited form as: Cell Rep. 2022 Feb 1;38(5):110312. doi: 10.1016/j.celrep.2022.110312

Figure 7. Zic5 stabilizes Gli3 through reducing its ubiquitination level.

Figure 7.

(A) Western blot analysis using the indicated antibodies on lysates from HEK293T cells transfected with control siRNA or Zic5 siRNA. Quantification of relative ratio of Gli3R/Gli3FL of western bolt with unpaired t test, ***P < 0.001. Histograms represent means ± s.d from three biological repeats.

(B) Zic5 stabilizes Gli3 in HEK293T cell by CHX assay. The relative levels of Gli3 full-length and repressor were quantified and normalized against α-tubulin from three independent experiments.

(C) Zic5 overexpression decreases Gli3 ubiquitination in HEK293T cell.

(D) Zic5 knockdown in HEK293T cells increases Gli3 ubiquitination.

(E) Zic5 knockdown in Xenopus eyes increases Gli3 ubiquitination.

(F) Cyclopamine treatment increases Gli3 ubiquitination and purmorphamine treatment decreases Gli3 ubiquitination in Xenopus eyes.

(G) Gli3/Gli3 interaction depends on 3rd-5th ZF domains.

(H) HEK293T cells were transfected with indicated plasmids and harvested 48h after transfection. Cells were lysed and immunoblotted with indicated antibodies.

(I) Gli3/Gli3 interaction increases its ubiquitination.

(G) HEK293T cells were transfected with indicated plasmids and harvested 48h after transfection. Cells were lysed and immunoblotted with indicated antibodies.