Movie 16.

Induction of ionic imbalance triggers LC3 association and prolongs SPN's intracellular survival. Time lapse fluorescence imaging of A549 cells stably expressing mStrawberry-Gal3 (red) and LC3-GFP (green) following infection with DRAQ5 stained Δply mutant SPN strain (magenta). Live imaging was performed under laser scanning confocal microscope starting at 30 min p.i. The video depicts association of Δply mutant SPN with LC3 without being associated with Gal3 following treatment with an ionophore, monensin. It also portrays prolonged persistence of SPN in these only LC3 marked non-canonical autophagosomes. Total 60 z-stacks were collected, out of which MIP of stacks 20-40 were shown in the movie. Time interval, 12 min; scale bar, 1.5 μm.