Tomograms from MEF cells (mitochondria)

Data includes ET tilt series and reconstructions from samples preprared on 17.1.2017 and 28.8.2013
Brendan Battersby, Paula Marttinen

2017
Cultured MEF cells were grown on glass coverslips and then fixed with 2% glutaraldehyde in Na-phosphate buffer, pH 7.4, for 30 min RT. After fixing, the samples were washed 3× 2 min with Na-phosphate buffer, pH 7.4. The samples were then post-fixed with 1% osmium tetroxide for 1 h at room temperature, dehydrated with a graded series of ethanol, incubated with transitional solvent acetone, and finally embedded gradually in epoxy. Dual axis tilt series were recorded from one or consecutive semi-thick (230-250 nm) sections using Tecnai FEG 20 microscope operating at 200 kV. The sections were tilted at one-degree intervals using a high-tilt specimen holder between ±62°. Images were acquired with SerialEM software (using a 4 k × 4 k Ultrascan 4000 CCD camera at nominal magnification of 9,600× (5 and 6) or 11,500× (1 and 2). The alignment of the tilt series and reconstructions were done with IMOD software package using 10-nm colloidal gold particles as fiducial markers.

#1 SC siRNA
#2 SFG3L2 siRNA
#5 MT-ATP6m.9205delTA SC siRNA
#6 AFG3L2 siRNA

Richter U, Ng KY, Suomi F, Marttinen P, Turunen T, Jackson C, Suomalainen A, Vihinen H, Jokitalo E, Nyman TA, Isokallio MA, Stewart JB, Mancini C, Brusco A, Seneca S, Lombès A, Taylor RW, Battersby BJ. Mitochondrial stress response triggered by defects in protein synthesis quality control. Life Sci Alliance. 2019 Jan 25;2(1):e201800219. doi: 10.26508/lsa.201800219. PMID: 30683687; PMCID: PMC6348486.
Published online 2019 Jan 25

Fig. 5B and C, videos 1 and 2

Battersby 2016: 28.8.2013, actinonin and Ctrl (EtOH)