FIGURE 5.

Lacritin-stimulated autophagy captures Htt103Q cellular aggregates. A, Htt103Q-mCFP or Htt25Q-mCFP constructs transduced in HCE-T cells stably expressing LC3 double tagged with EGFP and mCherry (22) (LC3/RG cells). B, quantitation of colocalized mCFP-Htt and mCherry LC3 at different times after treatment with 10 nm lacritin or C-25 in the absence of IFNG and TNF (t test; **, p < 0.01). C, number of LC3 mCherry- or LC3 mCherry-EGFP-labeled puncta in cells expressing Htt103Q or Htt25Q at different times after treatment with 10 nm lacritin or C-25 (no IFNG/TNF) (t test; *, p < 0.05). D, number of Htt103Q or Htt25Q puncta greater or less than 1 μm at different times after treatment with 10 nm lacritin or C-25 (no IFNG/TNF) (t test; *, p < 0.05; **, p < 0.01). Representative microscopic images are shown in E–H. E and F, LC3/RG cells expressing Htt103Q-mCFP that were treated with lacritin (E) or C-25 (F) for different times. Cells were washed and fixed, and all tags were localized. IFNG/TNF was omitted. G and H, LC3/RG cells expressing Htt25Q-mCFP that were treated with lacritin (G) or C-25 (H) for different times. IFNG/TNF was omitted. Bar, 10 μm. Error bars represent S.E. lacrt, lacritin.