Extended Data Fig. 1: Intracellular trafficking of pSTING.
From: Clathrin-associated AP-1 controls termination of STING signalling
a,b, Confocal imaging (a) or quantification of pSTING area in bright-field fluorescent microscopy images (b) of HeLaSTING cells stimulated for 0, 0.5, 1, 2, 3, 6, 9, 12 or 24 h with 1 µM diABZI. Cells were fixed and stained for TGN46 (trans-Golgi), pSTING and LAMP1 (endolysosomal compartment) as well as with Hoechst dye (nuclei, in dark blue). Mean ± s.e.m. of n = 3 independent experiments with at least 84 fields of view each per condition. Scale bars, 4 µm. c,d, Airyscan imaging (c) of HeLaSTING cells stimulated for 2.5 h with 1 µM diABZI. Cells were fixed and stained for TGN46 (trans-Golgi, not shown here), pSTING and the indicated marker as well as with Hoechst dye (nuclei, in dark blue). Colocalization of pSTING with the indicated marker is quantified by Manders’ colocalization coefficients (d). One representative cell is shown, and quantification is the mean ± s.e.m. of n = 8 cells examined in 1 of 4 independent experiments. Scale bars, 4 µm in larger left panel, 1 µm in zoomed-in panels. e, Airyscan imaging of HeLaSTING cells stimulated for 20, 150 (see Fig. 1b) or 360 min with 1 µM diABZI. Cells were fixed and stained for TGN46 (trans-Golgi), pSTING and CHC (clathrins) as well as with Hoechst dye (nuclei, in dark blue). One representative cell is shown of at least n = 6 cells from 3 independent experiments. White arrows point at occurrences of pSTING. Scale bars, 4 µm in larger left panel, 1 µm in zoomed-in panels.
