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. Author manuscript; available in PMC: 2021 Nov 9.
Published in final edited form as: Dev Cell. 2020 Sep 10;55(3):289–297.e4. doi: 10.1016/j.devcel.2020.08.008

Figure 2. Selective turnover occurs through an ATG5-regulated mechanism.

Figure 2.

(A) Lysosomes undergo rupture in response to LLOMe but not glucose starvation. Images show GFP-Gal3 fluorescence in LLOMe-treated cells at time zero after treatment (top left), or six hours after LLOMe washout (top right), and in control or cells starved of glucose for 24 hours (bottom images). (B) GFP-TRPML1 turnover in response to glucose starvation is regulated by ATG5 but not ATG13. Western blot shows GFP-TRPML1 cleavage after 24 hours of glucose starvation. GFP-TRPML1 and free GFP are indicated. Actin loading controls are also shown. (C) Quantification of data shown in part B from three independent experiments; error bars show SEM. (D) GFP-TRPML1 turnover in glucose-starved cells (24 hours) is not inhibited with treatment with the ULK1 kinase inhibitor SBI-0206965. Actin loading controls are also shown. (E) Autophagy flux in response to serum starvation for 10 hours, measured by GFP-LC3 cleavage leading to the generation of free GFP, is inhibited by treatment with SBI-0206965. Actin loading controls are also shown. (F) Quantification of data shown in part E from three independent experiments; error bars show SEM. *p<.05.