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. Author manuscript; available in PMC: 2021 Oct 1.
Published in final edited form as: J Cell Physiol. 2020 Mar 17;235(10):7309–7320. doi: 10.1002/jcp.29632

Fig. 4. 68 and Fx2149 increase ubiquitin-positive inclusions containing LRRK2.

Fig. 4.

A and B, HEK293T cells were transfected with WT- or Flag-G2019S-LRRK2 for 48 h, then treated with 0.2%DMSO, 68 (100nM), or Fx2149 (100 nM) for 24 h. Cells were subjected to immunostaining with anti-Flag (Green) and anti-ubiquitin (Red) antibodies. Merged images contain DAPI (blue) nuclear staining. A, Quantification data showed that both 68 and Fx2149 increased ubiquitin positive aggregates. P<0.05 by ANOVA, * vs cells expressing WT-LRRK2 treated with 0.2% DMSO; # vs vehicle treated control of each genotype cells. B. Representative images of LRRK2 transfected cells with ubiquitin positive aggregates. C and E, HEK293T cells were co-transfected with Flag-G2019S-LRRK2 and various HA-ubiquitin constructs for 48 h, and then treated with 0.2%DMSO, 68 (100nM), or Fx2149 (100 nM) for 24 h. Cells were immunostained with anti-Flag (green) and anti-HA(Red) antibodies along with DAPI (blue) nuclear marker. Transfected cells with LRRK2/ubiquitin positive aggregates were recorded. D. Representative images of co-transfected cells with aggregates. P<0.05 by ANOVA, * vs cells expressing G2019S-LRRK2 and wild type ubiquitin treated with 0.2% DMSO; # vs vehicle treated control cells with each ubiquitin construct group.