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The Ysa T3SS is necessary for intracellular replication within S2 cells. A gentamicin protection assay was performed using S2 cells seeded on glass coverslips. At the indicated times, samples were washed, fixed, and processed for fluorescence microscopy. Images were taken at a magnification of ×600. Green, gfp-expressing Y. enterocolitica; blue, nuclei stained with DAPI; S2 cells were visualized with differential interference contrast (DIC).
