Extended Data Fig. 6: Disruption of ER-TGN contact sites does not contribute to NLRP3 inflammasome activation.

a, Confocal images of HeLa cells expressing the ER-TGN membrane contact site reporters mCherry-FKBP-Cb5-mCherry and TGN46-FRB-HA treated with vehicle (Ctrl), 15 µM nigericin or 45 µg ml⁻¹ CL097 for 60 min. Cells were incubated with 200 nM rapamycin for 4 min before fixation. Cells were co-stained with antibodies against HA tag and GM130. Scale bar:10 µm. b, Quantification of cells showing co-localization of mCherry-FKBP-Cb5-mCherry and TGN46-FRB-HA in experiments shown in panel a. Cells were treated with 15 µM nigericin or 45 µg ml⁻¹ CL097 at indicated time points. Means of percentage ± SD, N = 4, n = 100 cells for each group. c, Immunoblotting of lysates from WT and ORP10 KO THP-1 cells. An antibody recognizing ORP10 was used. Tubulin was chosen as a loading control. d, Cellular uptake of Sytox Green in WT, VAP dKO and ORP10 KO (ORP10 KO-1) THP-1 cells treated with vehicle (Ctrl) (gray bars), 1 µg ml⁻¹ LPS (red bars) or 1 µg ml⁻¹ Pam3CSK4 (blue bars) for 2 hours. Means ± SD, N = 3. Data were analyzed with an unpaired two-sided t-test (b, d). Data shown in a, c are representative of three independent experiments.