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. 2022 Nov 28;24(1):30–41. doi: 10.1038/s41590-022-01355-3

Fig. 6. Defective ETT contributes to PI4P accumulation on endosomes.

Fig. 6

a, Confocal images of WT, ARFRP1 KO and SYS1 KO HeLa cells. Cells were co-stained with antibodies against TGN46 and giantin. DAPI was used to stain the nucleus. Scale bar, 10 μm. b, Quantification of percentage of cells with TGN46 on the Golgi in experiments shown in a. Mean of percentage ± s.d., N = 3, n = 100 cells for each group. c, Internalization of StxB in WT, ARFRP1 KO and SYS1 KO HeLa cells. Cells were fixed at 40 min after incubation with Cy3-conjugated StxB and stained with antibodies against EEA1 and giantin. DAPI was used to stain the nucleus. Scale bar, 10 μm. d, Quantification of cells with StxB on the Golgi in experiments shown in c. Mean of percentage ± s.d., N = 3, n = 100 cells for each group. e, Confocal images of WT, ARFRP1 KO and SYS1 KO HeLa cells expressing NLRP3-eGFP. Magnifications of areas in dashed squares are shown in the lower right corner. Arrowheads indicate EEA1-positive endosomes containing PI4P and NLRP3-eGFP. Scale bar, 10 µm. f, Quantification of NLRP3-eGFP puncta in experiments in a. Mean ± s.d., N = 3, n = 20 cells for each group. Data were analyzed with an unpaired two-sided t-test (b, d, f). Data shown in a, c and d are representative of at least three independent experiments.