Interplay between Leucine-Rich Repeat Kinase 2 (LRRK2) and p62/SQSTM-1 in Selective Autophagy
Fig 5
LRRK2 regulates Ser351 phosphorylation of p62.
A. Primary cortical neurons were infected with lentivirus that contain LRRK2 shRNA or a non-target shRNA sequence (NT shRNA) for 7 days, and endogenous protein expression levels were analyzed via western blotting with the indicated antibodies. B. Relative phosphorylation levels of p62 after LRRK2 knockdown normalized to NT shRNA were quantified. Summary data are presented as means ± SEM from three independent experiments (NT shRNA, 1.00 ± 0.00; pSer351, 2.92 ± 0.74; pSer403, 3.10 ± 1.09; n = 3; *p < 0.05, paired t-test). C. GFP-p62 WT or Ser351Glu (S351E) was co-transfected with vector control, WT, or G2019S myc-LRRK2 in HEK 293T cells. Endogenous Keap1 bindings to p62 were analyzed via immunoprecipitation with anti-GFP (p62) antibody. D. Quantification data from panel C represent means of relative Keap1 binding to p62 ± SEM (WT LRRK2, 1.05 ± 0.19; GS LRRK2, 0.67 ± 0.11; vector, 1.51 ± 0.08; n = 3; *p < 0.05, paired t-test).