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dc:title: Figure S3
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dc:description: <p>A) siRNA-treated cells were infected with <italic>C. trachomatis</italic> (MOI 1) for 24 h. Cells were then fixed and stained with an antibody specific for the Golgi protein GM130 (red channel) and chlamydial LPS (green channel). Samples were analysed at a LSCM. Overlays are shown. B) Expression of pmyc?Rab6AWT in Rab6A KD cells. At 26 h <italic>p.i.</italic>, cells were fixed and stained with antibodies specific for myc (red channel) and the Golgi protein Giantin (blue channel). Samples were analysed at a LSCM. Merge picture is shown. A single image for Giantin staining is shown in gray scale. C) Expression of peGFPRab11AWT in Rab11A KD cells. At 26 h <italic>p.i.</italic>, cells were fixed and stained with an antibody specific for the Golgi protein GM130 (red channel). Stable KD cells express eGFP as a marker for shRNA expression, therefore only high levels of eGFP-Rab11 could be detected. Intensity of the GFP channel was reduced to better visualize eGFP-Rab11. Samples were analysed at a LSCM. Merge pictures are shown. D) Rab1A-siRNA-treated Luciferase control cells were infected with <italic>C. trachomatis</italic> (MOI 1) for 24 h. Cells were then fixed and stained with an antibody specific for the Golgi protein GM130 (red channel) and chlamydial LPS (blue channel). Stable KD cells express GFP as a marker of shRNA expression. Samples were analysed at a LSCM. Overlays are shown. E) Golgin-84 cleavage in KD cells. Control (Luciferase), Rab6A KD and Rab11A KD cells were lysed in RIPA buffer and proteins were separated by SDS-PAGE. Proteins were transferred to PVDF membranes followed by immunoblot with antibodies specific for golgin-84 and actin. Actin was used as loading control. Images and blots are representative of n?=?3. Scale bar, 10 µm.</p>
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description: <p>A) siRNA-treated cells were infected with <italic>C. trachomatis</italic> (MOI 1) for 24 h. Cells were then fixed and stained with an antibody specific for the Golgi protein GM130 (red channel) and chlamydial LPS (green channel). Samples were analysed at a LSCM. Overlays are shown. B) Expression of pmyc?Rab6AWT in Rab6A KD cells. At 26 h <italic>p.i.</italic>, cells were fixed and stained with antibodies specific for myc (red channel) and the Golgi protein Giantin (blue channel). Samples were analysed at a LSCM. Merge picture is shown. A single image for Giantin staining is shown in gray scale. C) Expression of peGFPRab11AWT in Rab11A KD cells. At 26 h <italic>p.i.</italic>, cells were fixed and stained with an antibody specific for the Golgi protein GM130 (red channel). Stable KD cells express eGFP as a marker for shRNA expression, therefore only high levels of eGFP-Rab11 could be detected. Intensity of the GFP channel was reduced to better visualize eGFP-Rab11. Samples were analysed at a LSCM. Merge pictures are shown. D) Rab1A-siRNA-treated Luciferase control cells were infected with <italic>C. trachomatis</italic> (MOI 1) for 24 h. Cells were then fixed and stained with an antibody specific for the Golgi protein GM130 (red channel) and chlamydial LPS (blue channel). Stable KD cells express GFP as a marker of shRNA expression. Samples were analysed at a LSCM. Overlays are shown. E) Golgin-84 cleavage in KD cells. Control (Luciferase), Rab6A KD and Rab11A KD cells were lysed in RIPA buffer and proteins were separated by SDS-PAGE. Proteins were transferred to PVDF membranes followed by immunoblot with antibodies specific for golgin-84 and actin. Actin was used as loading control. Images and blots are representative of n?=?3. Scale bar, 10 µm.</p>
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dcterms:created: 2009-09-08T18:11:00
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title: Figure S3
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