Extended Data Fig. 10. Evaluation of the phosphoinositide-initiated membrane tethering and lipid transport pathway (PITT)-related markers PI4K2A and ORP9 in U2OS WT and U2OS G3BP double knockout cells after lysosomal damage.
a, b, Representative images of U2OS WT and U2OS G3BP DKO cells left untreated or treated with 2 mM LLOMe for 5 min, 45 min and 2 h, and stained for PI4K2A (a), ORP9 (b) and LAMP1(magenta). Note that quickly after LLOMe treatment PI4K2A and ORP9 localise to lysosomes. c,d, Shows high-content image quantification (n ≥ 300 cells) of PI4K2A (c) and ORP9 (d) puncta area per cell of cells treated as in (a,b). The graph on the right shows the corresponding co-localisation results at the indicated time points with the lysosomal marker LAMP1. Data represent the mean ± SEM. n ≥ 900 cells examined over three independent experiments. P-value was calculated using a two-way ANOVA, Šídák’s multiple comparisons test. Scale bar: 10 μm.