SEARCH

SEARCH BY CITATION

FilenameFormatSizeDescription
tra12158-sup-0001-FigureS1.docWord document256KFigure S1: Replicative vacuole assay in NLRC4−/− cells. NLRC4−/− bone marrow-derived macrophages were infected with L. pneumophila wild-type, ΔdotA and ΔsdcA-sidC strains for 1 h (A) or 8 h (B). Cells were fixed and stained with antibodies against ubiquitin (red) or Legionella (green). Nuclei were stained with Hoechst. C) Quantification of ubiquitin-positive vacuoles after 1 and 8 h. Average ± standard deviations were obtained from three independent experiments in which 300 vacuoles were scored for each strain. Statistical significance was obtained by comparing the number of cells showing ubiquitin-positive bacteria at 1 and 8 h between WT and ΔsdcA-sidC mutant (*p < 0.01).
tra12158-sup-0002-FigureS2.docWord document380KFigure S2: Single deletion mutants ΔsidC and ΔsdcA are not impaired for Arf1 recruitment. HEK293 cells stably expressing GFP-tagged Arf1 were infected with L. pneumophila wild-type, ΔsdcA, ΔsidC and ΔsidC_sdcA. A) Cells were fixed 1 h post-infection. Intracellular bacteria are shown in red. B) Quantification of the proportion of Arf1-positive vacuoles. Average ± standard deviations were obtained from three independent experiments (*p < 0.01).
tra12158-sup-0003-FigureS3.docWord document65KFigure S3: Rab1 is monoubiquitinated. HEK293 cells stably expressing Flag-tagged Rab1 were either uninfected or infected with L. pneumophila wild-type or ΔdotA for 1 h, following which they were lysed. FLAG-tagged Rab1 was immunoprecipitated using FLAG antibody-coated beads. Immunoprecipitated proteins were boiled and subjected to SDS–PAGE analysis and subsequently probed with either a Rab1A antibody or an ubiquitin-specific antibody FK2. The monoubiquitinated Rab1 band has been marked with an arrow in both blots.
tra12158-sup-0004-FigureS4.docWord document115KFigure S4: Dimeric SidCNT assembly found in the crystal. One SidCNT is shown in surface representation and the second SidCNT in ribbon representation.
tra12158-sup-0005-FigureS5.docWord document146KFigure S5: Analytical size exclusion chromatography of SidC shows that SidC is a monomer in solution. Analytical size exclusion chromatography (Superdex S75 10/30) of SidCFL, SidCΔC and SidCNT shows that SidC is a monomer in solution.

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.