Fig. 1: CHIP promotes the activation of Wnt signaling and the degradation of Arc. | Cell Death Discovery

Fig. 1: CHIP promotes the activation of Wnt signaling and the degradation of Arc.

From: CHIP promotes Wnt signaling and regulates Arc stability by recruiting and polyubiquitinating LEF1 or Arc

Fig. 1: CHIP promotes the activation of Wnt signaling and the degradation of Arc.

A CHIP increased the protein levels of β-catenin and non-phospho-β-catenin detecting by immunoblotting. HEK293T cells were transfected with an empty vector, Myc-CHIP, or Myc-CHIP T246M, and stimulated with Wnt3a (40 ng/ml) for 2 h. B HEK293T cells were transfected with an empty vector or HA-CHIP, and stimulated with Wnt3a CM or Ctrl-CM for 12 h. Wnt target genes (Myc, Cyclin D1, and Axin2) expression were analyzed by quantitative real-time PCR. C–D The knockdown efficiency of CHIP-specific siRNAs in protein level (C) and mRNA level (D). C, E Knockdown of CHIP reduced endogenous β-catenin detecting by immunoblotting (C), and inhibited Wnt target genes (Myc, Cyclin D1, and Axin2) expression detecting by quantitative real-time PCR (E) in SHSY5Y cells after CHIP-specific siRNAs transfection. F Western blot analysis of endogenous β-catenin in WT and CHIP−/− HT22 cells with Wnt3a stimulation or not. G–H Myc, Cyclin D1, and Axin2 mRNA abundance were analyzed by quantitative real-time PCR in WT and CHIP−/− HT22 cells (G), as well as WT and CHIP−/− 293T cells (H). I Immunoblot analysis of the protein level of β-catenin in cerebral cortex and cerebellum from the rat models of SCAR16 induced by CHIP T246M mutant or WT rats. J Immunoblotting analysis of the protein level of Arc after transfected with Myc-CHIP WT or Myc-CHIP T246M in SHSY5Y cells. All results were representative of three independent experiments. The graphs showed the means ± SD of three independent experiments. *P < 0.05, **P < 0.01, and ***P < 0.001 (Student’s t-test).

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