Biomedical Science Letters

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Fig. 1. The ubiquitination of LRRK2 wildtype and G2019S mutant are differentially regulated by p62/SQSTM1. (A) K63-linked ubiquitination is increased in LRRK2 G2019S mutant compared with LRRK2 WT. Myc-tagged LRRK2 WT or G2019S was co-transfected with HA-tagged K48 Ub or K63 Ub in HEK 293T cells. The ubiquitination of LRRK2 was detected using anti-HA antibody following immunoprecipitation with anti-myc antibody. (B) Quantification graph from panel A. The data represent means of ubiquitinated LRRK2 ± SEM (K48-Ub WT, 1.00±0.00; D1994A, 2.67±0.98; G2019S, 1.85±0.93; K63-Ub WT, 1.00±0.00; D1994A, 5.07±1.30; G2019S, 2.81±0.29; n = 3; *P < 0.05, paired t-test). (C) The overexpression of p62 reduces the ubiquitination of LRRK2 WT or D1994A, whereas does not have a significant effect on LRRK2 G2019S mutant. HEK 293T cells were co-transfected with HA-Ub and myc-LRRK2 WT or G2019S, and then the ubiquitination of LRRK2 was detected as in panel A. (D) The summary quantification are shown as means ± SEM (WT, 1.00±0.00; WT with p62, 0.48±0.18; D1994A, 0.69±0.06; D1994A with p62, 0.33±0.09; G2019S, 1.61±0.87; G2019S with p62, 0.60±0.32; n = 3; *P < 0.05, n.s., P > 0.05, paired t-test).
Biomed Sci Letters 2021;27:182-6
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