Fig. 2.

The role of ubiquitination of MHC II and CD86 by MARCH1 and MARCH8 in DC maturation. Surface expression of MHC II and CD86 in CpG-activated cDCs purified from the spleen of WT mice or mice deficient in either MARCH1 or MARCH8. Purified splenic cDCs (2x10⁵ ​cells) were kept on ice (-CpG) or incubated for 16 hours ex vivo with 50 nm CpG in 96-well plates, then washed and analyzed by flow cytometry for MHC II and CD86 surface expression. A fluorescence-minus-one (FMO) control was included, for which cells were incubated with the corresponding multi-colour staining panel, excluding the fluorescently labelled antibody species of interest (i.e. anti-CD86 or anti-MHC II mAb). Bars represent mean ​± ​SD with each symbol representing an individual mouse (n ​= ​6). Statistical analysis was performed using one-way ANOVA followed by Sidak's multiple comparisons test. ∗∗∗∗p ​< ​0.0001, ∗∗∗p ​< ​0.0002, ∗p ​< ​0.03.