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. 2017 Jun 21;37(25):5978–5995. doi: 10.1523/JNEUROSCI.3148-16.2017

Figure 6.

Figure 6.

UCH-L1 mainly deubiquitinates TrkB at K460 site. A, K460 is the major site of ubiquitination of TrkB. HEK293 cells were transiently transfected with Flag-TrkB FL or its site mutants together with Myc-UCH-L1 and HA-Ub. Cell lysates were immunoprecipitated using anti-Flag antibodies and the ubiquitination of TrkB was analyzed by immunoblotting with HA antibodies. B, Quantification of the ubiquitination level of WT or the site mutant TrkB FL. *p < 0.05, **p < 0.01 versus no UCH-L1 control, ##p < 0.01 versus WT TrkB-FL control, one-way ANOVA. C, UCH-L1 regulates TrkB ubiquitination at the K460 site. HEK293 cells were transfected with Flag-TrkB, Flag-TrkB KJMR, or Flag-TrkB R460K together with Myc-UCH-L1 and HA-Ub. Cell lysates were immunoprecipitated (IP) using anti-Flag antibodies and the level of ubiquitination of WT or mutant TrkB was analyzed by immunoblotting with HA antibodies. Statistical results are shown in D. **p < 0.01 versus no UCH-L1-expressing control, ##p < 0.01 versus WT TrkB-FL control, one-way ANOVA. All data are presented as mean ± SEM.