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. 2015 Mar 28;138(6):1531–1547. doi: 10.1093/brain/awv080

Figure 5.

Figure 5

CLEC16A is required for RILP-mediated perinuclear recruitment of HLA-II-positive late endosomes. (A) MelJuSo cells transfected with CLEC16A-GFP and RILP-RFP constructs were assessed for intracellular localization with HLA-DR by confocal microscopy. (B and C) GFP coimmunoprecipitations of HEK293T cells cotransfected with CLEC16A-GFP and (B) RILP-HA or (C) VPS41-HA constructs. GFP pull-downs were analysed for HA expression on immunoblots. Immunoprecipitated empty GFP (‘GFP’) constructs served as negative controls. These data are representative for 2–3 experiments. (D and E) MelJuSo cells treated with scrambled or CLEC16A siRNA were transfected with a RILP-GFP construct. (D) Intracellular localization (n = 3) and (E) surface expression (n = 3) of HLA-DR was evaluated in RILP-GFP+and GFP− cells 3 days after siRNA transfection [RILP-GFP+ versus GFP− scrambled siRNA-treated cells (scr): P = 0.06]. Scale bars = 5 μm.