Fig. 1.

Loss of VAMP8 results in decreased surface-levels of MHC class I. (A) siRNA knockdown for VAMP8 in human DCs (VAMP8 KD; NT: non-targeting siRNA; see reference (Dingjan et al., 2016) for quantification) and VAMP8 expression in BMDCs from VAMP8± and −/− mice by Western blot. GAPDH: loading control. (B) Representative flow cytometry histograms of surface staining of MHC class I and II molecules in VAMP8 KD (blue) and NT (green) human DCs (upper graphs) and for VAMP8−/− (blue) and VAMP8± (green) mouse BMDCs (lower graphs). The grey curves show the mean fluorescence intensities of the isotype controls. (C) Expression levels of HLA-A2 molecules (MHC-I), HLA-DR molecules (MHC-II), CD11c or CD86 on the surface of VAMP8 KD human DCs (left) or VAMP8-/- mouse BMDCs (right) normalized to control cells. (D) Representative dot blot cytokine array blots for NT and VAMP8 KD human DCs. The grey values are proportional to the levels of secreted cytokines. The 4 most abundant cytokines are indicated: IL-1ra; macrophage migration inhibitory factor (MIF); interleukin-8 (IL-8); Serpin E1. (E) Quantification of the cytokine secretion levels from panel D normalized to the highest intensities. Results are from at least three donors and plotted as mean ± s.e.m. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)