Fig 7.

A: Attachment of K. pneumoniae Ecl8, Ecl8ΔramA, Ecl8ΔramR or Ecl8ΔramRA to murine macrophage RAW 264.7 cell line. One-way ANOVA analyses were performed to demonstrate statistical significance. B: Microscopy to assess attachment to RAW 264.7 cell line. (i) Infection of the RAW264.7 cell line was carried out with K. pneumoniae Ecl8 (WT), Ecl8ΔramA, Ecl8ΔramR or Ecl8ΔramRA transformed with plasmid pRSMgfp. MOI was 1:100 and infections were carried out for 2 hrs. The actin cytoskeleton was stained with Acti stain 555 phalloidin (red) and host cell nuclei were stained with DAPI (blue). Images are representative of 80 fields. (ii) Graph representating mean values are derived from 3 independent experiments. One-way ANOVA analyses (P<0.001) were performed to demonstrate statistical significance. C: Internalisation of K. pneumoniae Ecl8, Ecl8ΔramA, Ecl8ΔramR or Ecl8ΔramRA by RAW 264.7 cells. Bacterial internalisation was assessed by the gentamicin protection assay. One-way ANOVA analyses were performed to demonstrate statistical significance. D: Enumeration of the extracellular non-phagocytosed K. pneumoniae Ecl8, Ecl8ΔramA, Ecl8ΔramR or Ecl8ΔramRA. One-way ANOVA analyses were performed to demonstrate statistical significance.