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Fig. 1. SNX1 is recruited to sites of Salmonella Typhimurium entry. HeLa cells were either fixed directly, or were infected with SL1344 for 15 min and fixed at indicated times. Cells were immunolabeled using anti-SNX1 (Alexa488, green) and stained with TRITC-phalloidin to visualize actin (red) and DAPI to label DNA (blue). (A) In uninfected cells, SNX1 displayed a cytoplasmic distribution with perinuclear enrichment. (B) SNX1 was recruited to invasion sites, readily identifiable by membrane ruffles, and (C) stayed associated with the bacteria. (D) After 180 minutes, SNX1 had resumed its original distribution. (E) For the kinetic analysis of SNX1 association with SCVs, between 50–150 SCVs were scored per assay per time point for the presence or absence of SNX1 (n
6 for 15 minutes and 60 minutes ± standard deviation, s.d.; n=2 for 180 minutes and 360 minutes, error ± minimum or maximum). (F) SNX1 recruitment to SCVs is comparable in MDCK (n=4, ± s.d., 50–150 SCVs scored per assay) and HeLa cells (n>3, ± s.d.), analyzed after a 15-minute bacterial `pulse' or further 45 minutes incubation. Scale bar: 10 µm.
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