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. 2023 Nov 15;7(2):e202302247. doi: 10.26508/lsa.202302247

Figure 8. DOCK1 promoted DUSP4 degradation via the ubiquitin proteasomal pathway.

Figure 8.

(A) DOCK1-knockout HTR-8 cells were transfected with HA-UB for 48 h and treated with 20 μM MG132 for 6 h. The cell lysates were immunoprecipitated with the DUSP4 antibody, and the ubiquitination assay showed the ubiquitination levels of DUSP4 protein. (B) HUWE1 protein levels after DOCK1 knockout or overexpression in HTR-8 cells. (C) After DOCK1 overexpression followed by HUWE1 knockdown, HTR-8 cells were treated with 50 μg/ml cycloheximide for 0, 1.5, 3, and 6 h, and Western blot was performed to detect DOCK1 and DUSP4 protein levels. (D) 293T cells were transfected with ectopic Flag-DOCK1, Myc-DUSP4, and HA-UB for 24 h, and then transfected with HUWE1 siRNA for 48 h. Finally, 20 μM MG132 was added and the protein was extracted after 6 h of incubation. The cell lysates were immunoprecipitated with Myc antibody and the ubiquitination assay showed DUSP4 protein ubiquitination levels.