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. 2014 Mar 17;204(6):909–917. doi: 10.1083/jcb.201308139

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© 2014 Ossareh-Nazari et al.

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Figure 1. — LTN1 deletion rescues the ribophagy defect of UBP3-null cells. (A–D) Cells expressing Rpl5-GFP (A and B) or Rpl25-GFP (C and D) were grown in rich medium (before starvation) and starved in SD-N for the indicated periods (starvation). (A and C, left) Degradation of GFP-tagged proteins was analyzed by anti-GFP blot of whole cell extracts. (right) The ratio between cleaved GFP and full-length protein was quantified for every time point in four independent experiments. a.u., arbitrary unit. (B and D) Cells before starvation or starved for 24 h (starvation) were examined both by fluorescence microscopy and differential interferential contrast (DIC). Note that 100% of wild-type (WT) and mutant cells expressed GFP-tagged proteins in the cytoplasm before starvation. C, cytoplasmic localization; V, vacuolar accumulation; C + V, localization in both cytoplasm and vacuole. Bars, 5 µm. The significance of the differences observed for the ribophagy efficiency was evaluated using Student’s t test. *, P = 0.01–0.05; **, P = 0.001–0.01. The errors bars correspond to standard deviations.