Supplementary Figure 2: Reversible block of ER marker proteins in autolysosomes obtained by cell exposure to the lysosomotropic drug BafA1.
From: Translocon component Sec62 acts in endoplasmic reticulum turnover during stress recovery
(a) After the CPA-induced stress, MEF were grown in the presence of BafA1 for 12 h and analyzed in IF (upper panels) or analyzed in IF 4 h after BafA1 wash out (lower panels). Scale bars, 10 μm. (b) Quantification of the Cnx-containing puncta in the upper and lower panels. n = 206 and 112 single cells for +BafA1 and BafA1 wash out, respectively. Mean ± SEM shown. ∗∗∗∗P < 0.0001, unpaired two-tailed t-test. Scale bars, 10 μm. (c) On inhibition of recovery from ER stress with BafA1 (+BafA1), Cnx accumulates in autolysosomes displaying GFP-Rab7 and Lamp1 in the limiting membrane. Scale bars, 10 μm. (d) Same as (c) for Sec62 (also refer to Fig. 2). Scale bars, 10 μm. Details of statistics in the Statistics and Reproducibility section. Statistics source data in Supplementary Table 4.
