Fig. 7: Pat1 is important for escape from the secondary vacuole.

a TEM images of WT and pat1::Tn mutant bacteria in HMECs at 48 hpi. “R” indicates R. parkeri and arrowheads point membranes surrounding the bacteria. Scale bar 1 µm. Images represent results from three independent experiments. b Percentage of bacteria in double-membrane compartments or in the cytosol (n = 3 independent experiments, >300 bacteria quantified), MOI 0.1. Data are mean ± SEM; **p = 0.0089 relative to WT (unpaired t-test (two-tailed)). c Percentage of WT and pat1::Tn mutant bacteria colocalizing with polyUb, p62, and NDP52 (all n = 4 independent experiments) at 48 hpi in HMECs (>1000 bacteria counted per strain/marker), MOI 0.5. Quantification was done using fluorescence microscopy with the same antibodies as Figs. 3a and 4a, b. Data are mean ± SEM; **p = 0.0017 (p62), p = 0.0067 (NDP52) relative to WT (unpaired t-test (two-tailed)). d Percentage of bacteria in the secondary cell that colocalize with the plasma membrane from the primary cell (measured using fluorescence microscopy) in mixed cell assays from Fig. 6c at 32 hpi (n = 3 independent experiments), MOI 4. Data represent individual fields imaged that contained bacteria in secondary cells (n = 46 fields for WT, n = 27 fields for pat1::Tn; bacterial number is in Fig. 6e). Data are mean ± SEM; **p = 0.0026 (Mann–Whitney (two-tailed)). Source data are provided as a Source Data file.