Figure 6: Pellino-mediated MyD88 ubiquitination is dependent on CTE domain.
(a,b) S2 cells were transfected with Myc-MyD88ΔCTE and HA-Ub with Flag-Pellino (a), or also pretreated with pellino dsRNA (b). Cell lysates were used to perform immunoprecipitation experiments with anti-Myc beads followed by immunoblot analysis with anti-HA antibodies to show ubiquitination patterns of MyD88ΔCTE. Expression levels of transfected proteins and Pellino knockdown efficiency in whole-cell lysates are shown in the bottom panel. (c,d) S2 cells transfected with Flag-Pellino alone or in combination with Myc-tagged MyD88 CTE domain were stained with Hoechst and anti-Flag antibody alone or with anti-Myc antibody as indicated and imaged by confocal microscopy (c). Scale bars, 10 μm. (d) Statistical assays of PM or CP localization of Pellino in transfected cells in panel (c). (e,f) S2 cells transfected with Flag-Tube alone or in combination with Myc-tagged MyD88 or MyD88 CTE domain were stained with Hoechst and anti-Flag antibody alone or with anti-Myc antibody as indicated and imaged by confocal microscopy (e). Scale bars, 10 μm. (f) Quantification assays of P.M. or C.P. localization of Tube in transfected cells in (e).
