Figure 4.
IST1 structures are devoid of phosphoinositide (PI) lipids. (A) Representative TIRF microscopy of moDCs co-expressing mCherry-tagged CHMP4 (magenta in merge) with GFP-tagged probes for PI lipids. The following PI probes were used: the PH-domain of PLCδ1 for PI(4,5)P2, the PH-domain of AKT for PI(3,4,5)P3, the N-terminal sequence of MCOLN1 for PI(3,5)P2, and the PX-domain of NCF4 (p40phox) for PI(3)P (n = 3 donors). The line graphs show fluorescence intensity profiles as indicated by the yellow lines. The yellow shaded areas indicate regions where the fluorescence intensity of both proteins exceeds the 50% intensity. (B) Confocal micrographs of monocyte-derived macrophages (MΦ), CD1c+ dendritic cells, primary dermal fibroblasts, and mouse macrophage cell line RAW 264.1. Arrows indicate IST1 structures. Scale bars: 10 μm.

IST1 structures are devoid of phosphoinositide (PI) lipids. (A) Representative TIRF microscopy of moDCs co-expressing mCherry-tagged CHMP4 (magenta in merge) with GFP-tagged probes for PI lipids. The following PI probes were used: the PH-domain of PLCδ1 for PI(4,5)P2, the PH-domain of AKT for PI(3,4,5)P3, the N-terminal sequence of MCOLN1 for PI(3,5)P2, and the PX-domain of NCF4 (p40phox) for PI(3)P (n = 3 donors). The line graphs show fluorescence intensity profiles as indicated by the yellow lines. The yellow shaded areas indicate regions where the fluorescence intensity of both proteins exceeds the 50% intensity. (B) Confocal micrographs of monocyte-derived macrophages (MΦ), CD1c+ dendritic cells, primary dermal fibroblasts, and mouse macrophage cell line RAW 264.1. Arrows indicate IST1 structures. Scale bars: 10 μm.

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