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. 2011 Apr 25;108(19):7826–7831. doi: 10.1073/pnas.1013800108

Fig. 5.

Fig. 5.

spin is required for mTOR reactivation following starvation. (A) NRK-LC3 cells were transfected with nonspecific RNAi (NS) or RNAi against spin. Sixty hours after transfection, cells were starved 0, 2, 6, or 8 h, harvested, and analyzed by Western blot using antibodies against Phospho-p70 S6 Kinase (P-S6K), Total S6 Kinase (S6K), and Actin. (B and C) NRK cells were transfected with spin-RNAi as in A, and starved for 10 h. FCS was then added back for 0–120 min; cells were harvested and analyzed by Western blot using antibodies against Phospho-p70 S6 Kinase (P-S6K) (B), or observed by confocal microscopy for the Lamp1 intracellular pattern (C). (Scale bar, 5 μm.) (Inset) Enlargement of tubules extruding from lysosomes.