Fig. 5: Pat1 is important for avoiding bacterial association with damaged membranes.

a Images of Gal3-mCherry (magenta in merge) and autophagy receptors p62 (left; cyan in merge) and NDP52 (right; cyan in merge) in HMECs infected with WT or pat1::Tn bacteria (green in merge) at 1 hpi. Arrows indicate large Gal3 positive clusters near bacteria, asterisk indicates bacteria that are p62 positive and Gal3 negative, arrowheads indicate colocalization between NDP52, Gal3, and bacteria. b Images of Gal8-mCherry (magenta in merge) and autophagy receptors p62 (left; cyan in merge) and NDP52 (right; cyan in merge) in HMECs infected with WT or pat1::Tn bacteria (green in merge) at 1 hpi. Arrows indicate large Gal8 positive clusters near bacteria, asterisk indicates bacteria that are p62 positive and Gal8 negative, arrowheads indicate colocalization between NDP52, Gal8, and bacteria. Scale bar for (a) 5 µm, (b) 3 µm. c Quantification of (a), percentage of bacteria positive for Gal3 (>500 bacteria counted per strain), MOI 5. d Quantification of (b), percentage of bacteria positive for Gal8 (>500 bacteria counted per strain), MOI 5. Data in (c, d) are mean ± SEM; ***p = 0.0008 **p = 0.0020 relative to WT (unpaired t-test (two-tailed)). e Quantification of (a) and (b), percent of polyUb, p62, or NDP52 positive pat1::Tn bacteria that are also positive for Gal3 (open diamond) or Gal8 (filled diamond)(50–60 polyUb, p62, or NDP52 positive bacteria in Gal+ cells were counted for both Gal3 and Gal8), MOI 5. f Percent of bacteria positive for NDP52 in untreated cells or cells that undergo hypotonic lysis of vesicles (>1000 bacteria counted per strain/condition), MOI 5. Data are mean ± SEM; **p = 0.0063 relative to untreated (paired t-test (two-tailed)). c, d Data represent n = 4 independent experiments, e, f data represent n = 3 independent experiments. Source data are provided as a Source Data file.