2024-06-26 REL PDBe PDBe PDBe 2024-04-10 2024-06-19 2024-06-19 2024-06-26 Vienna Science and Technology Fund (WWTF) LS19-001 Austria Austrian Science Fund P 36743-B Austria Bragulat-Teixidor H Otsuka S Endoplasmic reticulum, Nuclear envelope, Organelle contact site, Membrane junction, CELL CYCLE Bragulat-Teixidor H Ishihara K Szucs GM Otsuka S Embo Rep. UK 2024 38877171 doi:10.1038/s44319-024-00175-w 1469-3178 HeLa Kyoto cell HeLa Kyoto cell 0 Homo sapiens HeLa tomography cell 7.4

DMEM without Riboflavin and Phenol Red, containing 10% FBS, 1% Pen/Strep, and 50 nM SiR-DNA

NEGATIVE Uranyl acetate and lead citrate Agar 100 Epoxy resin

Frozen cells were substituted in 0.1% uranyl acetate (UA), 2% Osmium tetroxide and 5% H2O in acetone following this temperature ramp: -90 C to -80 C for 10 hours, -80 C to -30 C for 10 hours, -30 C for 4 hours, -30 C to 0 C for 6 hours, 0 C to 20 C for 4 hours, 20 C for 5-6 hours. Afterwards, samples were washed three times in pure acetone for at least 10 minutes each, and subsequently infiltrated with Agar 100 Epoxy resin. The resin infiltration was done progressively at room temperature with increasing concentrations of resin in acetone (3:1 for 2-3 hours, 1:1 for 2-3 hours, and 1:3 overnight). Infiltration with pure resin was done at room temperature for at least 5 hours. Resin was polymerized at 60 C for 72 hours.

NITROGEN Cytodiagnostics 15 OTHER

High pressure freezing chamber was 1 mm thick, 6.0 mm diameter, with central cavities 5.0 mm x 5.0 mm x 25 um deep. The chamber had been in contact with 1-hexadecene.. The value given for _em_high_pressure_freezing.instrument is Leica EM ICE. This is not in a list of allowed values {'OTHER', 'LEICA EM PACT', 'LEICA EM HPM100', 'EMS-002 RAPID IMMERSION FREEZER', 'BAL-TEC HPM 010', 'LEICA EM PACT2'} so OTHER is written into the XML file.

20% Ficoll-PM400 Leica Ultracut UCT 298 250 FEI TECNAI F20 FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 200 0.2 0.5 FEI EAGLE (4k x 4k) 40.0 1 BACK PROJECTION 110