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. 2019 Mar 4;15(8):1333–1355. doi: 10.1080/15548627.2019.1581009

Figure 7.

Figure 7.

The ATG14 LIR motif is important for binding of the PtdIns3K-C1 complex to GABARAP and GABARAPL1, and for activation of the complex by ULK1-mediated Ser29 phosphorylation of ATG14. (a) GST affinity isolation using bead-bound GST-Atg8-family fusion proteins and lysates from HCT116 cells knocked out for ATG14 and reconstituted with WT or LIR mutated MYC-ATG14. Bead bound proteins were detected by western blots using the indicated antibodies. (b) MYC-Trap experiments performed with the indicated cell lines. Bound proteins were detected by western blots using the denoted antibodies. (c) MYC-Trap experiments performed with the indicated cell lines, incubated in starvation medium (HBSS) for 6 h or not. Bound proteins were detected by western blots using the indicated antibodies. (d-g) Relative quantifications of the data in (c), showing the relative amount of BECN1, p-S93 BECN1 and p-S29 MYC-ATG14 in the MYC precipitates. Data information: Means ± SD of 3 independent experiments. Significant P values are indicated (Student´s two-tailed, unpaired t-test). ns, not significant.