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. 2018 Sep 10;131(17):jcs211672. doi: 10.1242/jcs.211672

Fig. 1.

Fig. 1.

The PX domain of SNX21 is required for its targeting to highly dynamic PtdIns(3)P-enriched early endosomes. (A) HeLa cells stably expressing a plasmid encoding eGFP-SNX21 were imaged live. A selected frame of a live movie depicting the localisation of GFP-SNX21 to highly dynamic, peripherally localised punctae. Scale bar: 40 µm. (B) Protein sequence alignments between SNX1, SNX3 and SNX21 reveal the conserved arginine residues in the PX domain of SNX21 implicated in the binding to phosphoinositides. (C) HeLa cells were transfected with DNA encoding eGFP-SNX21, fixed and imaged: wild-type SNX21 localises to peripheral punctae, SNX21 R171A is cytosolic, as is wild-type SNX21 upon inactivation of PI3-kinase via treatment with wortmannin (200 nM). Scale bars: 20 µm. (D) HeLa cells were virally transduced to express GFP-SNX21 and co-immunostained for endogenous proteins representative of various trafficking compartments and imaged using confocal microscopy. Scale bars: 20 µm. (E) Quantitative colocalisation analysis between GFP-SNX21 and endogenous compartment markers. Graph represents the mean of >22 cells quantified; error bars show s.e.m.