Fig. 4.

AP-3 fails to be recruited to TLR9⁺ endosomes after CpG stimulation in the presence of PIKfyve inhibitor. RAW 264.7 cells stably transduced with TLR9-GFP and AP-3μ3A-Flag were stimulated with DOTAP-CpG-A-Cy5 for 1 h in the absence (C) or presence (D) of YM201636 (1 μM) given 1 h prior to CpG-A stimulation. Subsequently, cells were stained and analyzed by confocal microscopy to detect TLR9-GFP (green), AP-3 (red) and CpG (blue). Scale bars, 2.5 μm. No-stimulation controls in the absence or presence of inhibitor are shown as (A), (B), respectively. ***P < 0.001.