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. 2016 Sep 14;5(10):1516–1529. doi: 10.1242/bio.018648

Fig. 5.

Fig. 5.

Localization of STX17 is affected in the absence of LAMP-2. In LAMP-2-negative fibroblasts, STX17 could not be detected on either LC3-positive autophagosomes (A) or on LAMP-1-positive vesicles (B). In wild-type MEFs, STX17 is mainly present on the autophagosomes as indicated by co-localization with LC3 (C) and occasionally on LAMP-1-positive lysosomes (D). Scale bars=20 µm. Using the JACoP plugin, a significant decrease of co-localization of STX17 with LC3 was observed in the absence of LAMP-2 (E), while the overlapping of STX17 with LAMP-1 was absent in both cell types (F). Immuno-electron microscopy confirms the autophagosomal localization of STX17 (15-nm gold particle, arrowheads) in wild-type MEFs (G). Due to the low number of STX17-positive particles definitive localization could not be established in LAMP-2-single- and LAMP-1/2-double-deficient cells. Scale bars=500 nm. (G). LAMP-1 and STX17 were observed in independent vesicles in wild-type MEFs. Scale bars=200 nm (H). Data are expressed as mean±s.d. from at least 15 cells in each condition and are representative of three independent experiments. **P<0.01; ****P<0.0001 (Mann–Whitney test). L, lysosomes; AP, autophagosome.