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Fig. 3. Effect of DAT on phosphorylation and polyubiquitination of APPL1 in HepG2 cells. (A) Impact of DAT on the levels of intracellular APPL1 protein. (B) Impact of DAT on serine phosphorylation of APPL1. APPL1 was immunoprecipitated by specific antibody and serine phosphorylation was then detected by western blot using a specific anti-serine antibody. (C) Impact of DAT on K63-linked polyubiquitination of APPL1. APPL1 was immunoprecipitated by specific antibody and polyubiquitination was detected by Western blot using a specific antibody against K63-linked polyubiquitination. (D) Quantitative analysis of K63-linked polyubiquitination of APPL1 in (C). (E) Impact of DAT on the levels of intracellular Nedd4 and TRAF6 proteins. (F) Quantitative analysis of TRAF6 levels in (E). (G) Impact of TRAF6 knockdown on K63-linked polyubiquitination of APPL1. DAT, diallyl trisulfide; APPL1, adaptor protein, phosphotyrosine interacting with PH domain and leucine zipper 1; K63, Lys 63; Nedd4, neural precursor cell expressed, developmentally down-regulated protein 4; TRAF6, TNF receptor-associated factor 6; IP, immunoprecipitation; Ub, ubiquitin; polyub, APPL1 polyubiquitination. *p < 0.05, **p < 0.01 vs. control group.
Korean J Physiol Pharmacol 2022;26:457-468 https://doi.org/10.4196/kjpp.2022.26.6.457
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