Data includes ET tilt series and reconstructions from samples prepared on 2.10.2009. SBF-SEM data set from sample 20.5.2012, run 30.6.2012. Eeva-Liisa Eskelinen, Joanna Biazik NRK-52E cells were grown on 3-cm dishes containing glass cover slips to subconfluency. The cells were fixed in 2% GA in 0.1 M sodium cacodylate buffer, pH 7.4, postfixed in reduced osmium tetroxide, and flat embedded in Epon. Semithick 250-nm sections were prepared and picked up on single slot grids. Colloidal gold particles, 10 nm in diameter, were placed on top of the sections to serve as fiducial markers for alignment of the tomograms. Dual axis tilt series were acquired using SerialEM software running on a Tecnai FEG 20 microscope operating at 200 kV with a nominal magnification of 19 000 x giving a final pixel size of 0.56 nm. Images were recorded at 1 degree intervals for the a-axis and 1.5 degree intervals for the b-axis with either a 1 k × 1 k or a 4 k × 4 k CCD camera over a tilt range of ±62 degrees. IMOD software was used to create 3D reconstructions from the tilt series and to create 3D models of the membranes. Tilt series were captured from 6 regions (from different cells) and segmentation and 3D modeling was performed on 3 of those tomographic sequences. Biazik J, Ylä-Anttila P, Vihinen H, Jokitalo E, Eskelinen EL. Ultrastructural relationship of the phagophore with surrounding organelles. Autophagy. 2015;11(3):439-51. doi: 10.1080/15548627.2015.1017178. PMID: 25714487; PMCID: PMC4502653. Figs. 4 and 5.

2015