Immediately ex-vivo NOD Th40 cells have CD40 and TRAF2 associated with rafts.

Sorted Th40 cells and CD4^hi T cells from age matched 8–12 week-old female NOD (euglycemic), BALB/c or NOR were immediately lysed (Imm. ex-vivo) or untreated (UN), CD40-crosslinked (CD40), Fas-crosslinked (Fas) or (CD40+Fas)-crosslinked ((CD40+Fas)) for 5 hours then soluble (S) and insoluble (I) fractions were prepared. CD40 levels in Th40 cells and CD4^hi (A and B respectively) and TRAF2 levels in Th40 cells and CD4^hi (C and D respectively) were analyzed in western blots. CD40 was analyzed first then the membranes were stripped and TRAF2 was analyzed. As internal loading standard (std) membranes were stripped and stained with coomassie blue and a representative band of the same size from each membrane is shown. Data are representative of 3 separate experiments.