Fig. 4: CHIP increases the K63-linked ubiquitination of LEF1 and the K48-linked ubiquitination of Arc. | Cell Death Discovery

Fig. 4: CHIP increases the K63-linked ubiquitination of LEF1 and the K48-linked ubiquitination of Arc.

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

Fig. 4: CHIP increases the K63-linked ubiquitination of LEF1 and the K48-linked ubiquitination of Arc.

A CHIP promoted the ubiquitination of LEF1. Immunoprecipitation and immunoblot analysis were performed to detect the ubiquitination of LEF1 after co-transfection of HA-tagged ubiquitin, Flag-LEF1, and Myc-CHIP in HEK293T cells. B Knockdown of CHIP reduced the ubiquitination of LEF1. HEK293T cells were transfected with HA-tagged ubiquitin, Flag-LEF1, and CHIP-specific siRNA or control siRNA (NC) (40 nM), then the cell lysates were detected by co-immunoprecipitation and immunoblot. C–E HEK293T cells were transfected with indicated plasmids or CHIP-specific siRNA as figures shown. Whole cell extracts were immunoprecipitated with anti-Flag beads and blotted with anti-HA antibody. The inactive mutants of CHIP couldn’t enhance the ubiquitination of LEF1 as CHIP WT (C). CHIP increased the K63-linked ubiquitination of LEF1 (D). Knockdown of CHIP reduced the K63-linked ubiquitination of LEF1 (E). F–H Immunoprecipitation and immunoblot analysis were performed to detect the ubiquitination of Arc after co-transfection with indicated plasmids or CHIP-specific siRNA as figures shown. The inactive mutants of CHIP couldn’t enhance the ubiquitination of Arc (F). CHIP increased the K48-linked and K63-linked ubiquitination of Arc (G). Knockdown of CHIP reduced the ubiquitination, even the K48-linked ubiquitination of Arc (H). All results were representative of three independent experiments.

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