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. Author manuscript; available in PMC: 2010 Aug 18.
Published in final edited form as: Cell Host Microbe. 2010 Feb 18;7(2):115–127. doi: 10.1016/j.chom.2010.01.007

Figure 1. SIN Induces Autophagy in vitro.

Figure 1

(A) Quantitation of the percentage of GFP-LC3 MEFs with GFP-LC3 punctae (autophagosomes) after infection with indicated virus. Data shown represent mean ± SEM for triplicate samples of at least 100 cells per sample. Similar results were observed in 3 independent experiments.

(B) Representative fluorescent microscopic image demonstrating colocalization of SIN structural proteins (red) with GFP-LC3 (green) in GFP-LC3 MEFs at 12 h post-infection (p.i.).

(C) Representative fluorescent microscopic images showing colocalization in Atg5+/+ MEFs or lack of colocalization in Atg5−/− MEFs of SIN capsid (red) and GFP-LC3 (green) in cells infected with SIN-mCherry.capsid/GFP-LC3. Images shown represent a single time point at 12 h p.i. from live cell imaging. Arrowhead denotes colocalized puncta; arrow denotes capsid-positive GFP-LC3 ring structure. See movie S1 for dynamic representation of mCherry.capsid and GFP-LC3 localization between 16 and 17 h p.i. in Atg5+/+ MEFs.

(D) Representative EMs of wild-type MEFs at 12 h p.i. with SVIA. Left panel demonstrates a double-membraned autophagosome (black arrow) containing SIN nucleocapsids (black arrowheads), cellular membranes (white arrowhead), and aggregates (white arrow). Right panel demonstrates a single-membraned autolysosome with SIN nucleocapsid (black arrowhead). Open arrowheads denote virions budding from the plasma membrane. Scale bars, 200 nm.

(E) Measurement of autophagic protein degradation by p62 Western blot analysis in wild-type MEFs at serial time points after mock, SVIA, or UV-SVIA infection.