Figure 5: Toll signalling promotes Pellino to accumulate at the plasma membrane in a MyD88-dependent manner.
(a) S2 cells were transfected with Myc-tagged Pellino or also pretreated with MyD88 or Tube dsRNA, and then treated with or without pre-boiled M. luteus (ML) for 12 h as indicated. Cells were further stained with anti-Myc antibody and Hoechst, and imaged by confocal microscopy. Scale bars, 10 μm. (b) Statistical assays of plasma membrane (PM) or cytoplasmic (CP) localization of Pellino in cell samples from (a). (c,d) S2 cells transfected with Myc-Pellino alone or Myc-Pellino in combination with Flag-MyD88 were stained with Hoechst and anti-Myc antibody alone or with anti-Flag antibody as indicated and imaged by confocal microscopy(c). Scale bars, 10 μm. (d) Statistical assays of PM or CP localization of Pellino in cell samples from (c). (e) Schematic diagram of MyD88 and its truncated mutants. (f) S2 cells were transfected with Flag-tagged Pellino and Myc-tagged MyD88 or its truncated mutants. Forty-eight hours after transfection, cell lysates were prepared, immunoprecipitated with anti-Myc beads, followed by immunoblot analysis with the indicated antibodies. (g,h) S2 cells transfected with Flag-Pellino alone or in combination with Myc-MyD88ΔCTE and stained with Hoechst and anti-Flag antibody alone or with anti-Myc antibody as indicated and imaged by confocal microscopy (g). Scale bars, 10 μm. (h) Statistical assays of PM or CP localization of Pellino in cell samples from (g).
