Figure 6. Increased V0–V1 engagement is... | Rockefeller University Press

Figure 6.

$Increased V0–V1 engagement is associated with activation of non-canonical autophagy. (a) WT MCF10A cells were treated with BafA1 (100 nM), SaliP (2.5 μM), monensin (100 μM), or CQ (100 μM) for 1 h. Following fractionation, input, membrane, and cytosol fractions were probed for ATP6V1A, ATP6V0D1, and LC3 by Western blotting. LC3II/LC3I ratios are shown below. (b) Quantification of V0–V1 association from experiments as shown in a. Data represent mean ± SD from four to seven independent experiments. ***, P < 0.003; **, P < 0.01; *, P < 0.04, unpaired t test. (c) HeLa cells were stimulated with either mTOR inhibitor AZD8055 (1 μM) or TRPML1 agonist C8 (2 μM) for 90 min. Following fractionation, membrane and cytosol fractions were probed for ATP6V1B2, ATP6V0D1 by Western blotting. (d) Quantification of V0–V1 association from experiments as shown in c. Data represent mean ± SD from two independent experiments. Source data are available for this figure: SourceData F6.$

Increased V0–V1 engagement is associated with activation of non-canonical autophagy. (a) WT MCF10A cells were treated with BafA1 (100 nM), SaliP (2.5 μM), monensin (100 μM), or CQ (100 μM) for 1 h. Following fractionation, input, membrane, and cytosol fractions were probed for ATP6V1A, ATP6V0D1, and LC3 by Western blotting. LC3II/LC3I ratios are shown below. (b) Quantification of V0–V1 association from experiments as shown in a. Data represent mean ± SD from four to seven independent experiments. ***, P < 0.003; **, P < 0.01; *, P < 0.04, unpaired t test. (c) HeLa cells were stimulated with either mTOR inhibitor AZD8055 (1 μM) or TRPML1 agonist C8 (2 μM) for 90 min. Following fractionation, membrane and cytosol fractions were probed for ATP6V1B2, ATP6V0D1 by Western blotting. (d) Quantification of V0–V1 association from experiments as shown in c. Data represent mean ± SD from two independent experiments. Source data are available for this figure: SourceData F6.