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. Author manuscript; available in PMC: 2019 Jul 17.
Published in final edited form as: Sci Transl Med. 2018 Jun 6;10(444):eaan8162. doi: 10.1126/scitranslmed.aan8162

Fig. 4. LRRK2 augments Beclin-1 degradation blocking autophagy and increasing LRRK2 expression.

Fig. 4.

(A) BMDCs from Lrrk2 Tg mice or littermate control mice were stimulated for 6 hours with M. leprae. The cell lysates were then subjected to immunoblotting with the anti–Beclin-1 antibody H-300. (B) Whole-cell lysates of HEK293T cells cotransfected with the indicated plasmids were sonicated and subjected to immunoprecipitation with anti-FLAG antibody, followed by immunoblotting with anti-HA antibody. Immunoblotting using anti–Beclin-1 antibody (clone: 20/Beclin) demonstrated degradation bands (black arrows). (C and D) LRRK2 mRNA and protein expression was determined using real-time PCR (C) and immunoblotting (D) in HCT116 WT cells, Beclin-1 KO cells, and ATG5 KO cells. Data are representative of at least three identical experiments. (C) HCT116 WT versus Beclin-1 KO, P = 0.0009; HCT116 WT versus ATG5 KO, P = 0.0015. *P < 0.05 was considered a statistically significant difference. Statistical significance was determined by analysis of variance (ANOVA) for multiple comparisons.