Fig. 6.

Atg40 and Atg11 fail to colocalize in the lnp1∆ mutant and in LAT-A–treated cells during the induction of ER-phagy with rapamycin. (A) WT and lnp1Δ cells expressing Atg40-3xGFP and Atg11-2xmCherry were incubated for 3 h at 30 °C in SC media containing rapamycin (200 ng/mL) before they were directly examined by fluorescence microscopy. Arrowheads point to Atg11 puncta colocalized with Atg40 puncta. (A, Right) Quantitation of the percent of Atg11-2xmCherry puncta colocalized with Atg40-3xGFP puncta after 0 or 3 h of rapamycin treatment. Error bars represent SEM from three separate experiments. **P < 0.01, Student’s t test. (B) Same as A, only WT cells were treated for 3 h with rapamycin in the absence or presence of 100 µM LAT-A. Arrowheads point to Atg11 puncta colocalized with Atg40 puncta. (B, Right) Quantitation of the percent of Atg11-2xmCherry puncta colocalized with Atg40-3xGFP puncta after 0 or 3 h of rapamycin treatment. Error bars represent SEM from three separate experiments. ***P < 0.001, Student’s t test. (Scale bars, 2 μm.)