Fig. 4

ATG14-associated Vps34 activity is reduced in Q175 brains. a ATG14 phosphorylation in Q175 brains. ATG14 was immunopurified using antibodies against ATG14. Two representative lanes shown for each experiment. b Quantification of ATG14 phosphorylation. **p < 0.01 (n = 5). Data are represented as mean +/− SEM. c In-vitro lipid kinase assay of Vps34 complex pulled down with antibodies against ATG14 from the brains of Q175 mice. Thin layer chromatography was used to separate ³²P-PI(3)P. d Quantification of PI(3)P levels were normalized to levels of Vps34 in the IP. ***p < 0.001 (n = 4). Data are represented as mean +/− SEM. e Beclin 1 S14 phosphorylation in Q175 mice. f Quantification of Beclin 1 phosphorylation. **p < 0.01 (n = 5). Data are represented as mean +/− SEM. g ATG14 phosphorylation in mCFP-103Q expressing HeLa cells. Doxycycline was removed for 7 days to induce protein expression. IP was performed against ATG14. h Quantification of ATG14 phosphorylation. Values were normalized to control. *p < 0.05 (n = 4). Data are represented as mean +/− SEM. I Immunofluorecent imaging of autophagosome reporter GFP-LC3 distribution in striatal nuclei of Q175;GFP-LC3 and control WT;GFP-LC3 mice. Mice at 6, 10 and 15 months are examined. mHtt nuclear aggregate staining in red and GFP-LC3 in green. Scale bar 10 μm