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. 2014 Nov 3;124(12):5249–5262. doi: 10.1172/JCI77598

Figure 5. A failure of clearance of autophagosomes in spastizin- or spatacsin-depleted cells occurs downstream of mTOR activation.

Figure 5

(A and B) HeLa cells were transfected with siCTL, siSPG15, or siSPG11 siRNAs; starved by serum deprivation; and immunostained for LC3. (A) Representative images of starved control cells for the indicated time periods. (B) Starved cells with >20 autophagosomes were quantified (n = 3; >200 cells per experiment). (C) Lysates from A were immunoblotted with the indicated antibodies. Molecular weight standards (in kDa) are shown to the left. Molecular weight of S6K is approximately 59 kDa. (D) Phospho-S6K levels from C were normalized to total S6K (n = 3). (E) HeLa cells were transfected with siCTL, siSPG15, or siSPG11 siRNAs and then coimmunostained for mTOR (green) and LAMP1 (red). Scale bar: 10 μm. Mean ± SD are shown.