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Cortical neurons, fibroblasts and astroglial cells from Ophn1 knock-out animals and wild-type controls in C57BL/6 background were prepared by standard procedures as previously described (7).
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For this purpose, a suspension of 100 µg/ml with a 1 mM KNO3 background was prepared by sonication for three minutes with 80%% amplitude (Disintegrator UDS 751 equipped with a 14 mm titanium probe tip, Topas GmbH, Dresden, Germany).
Background control wells were prepared by omitting primary antibodies, DRAQ5 and Sapphire700 (i.e. secondary only).
Background electrolytes (BGEs) containing proteins were prepared by dissolving GST-fused Apm1 or GST-fused Rho3 (as a negative control) at a concentration of 200 µg/ml or 100 µg/ml.
Over 500,000 negative samples were prepared by randomly cropping 1,085 background images that did not contain any vehicle objects.
RORγt Tg mice were prepared by backcrossing mice on the C57BL/6 background.
Transgenic flies were prepared by P-element-mediated transformation in w 1118 background.
Progenies used in aging assays and metabolite profiling experiments were prepared by mating wild-type animals of Canton-S background, which were backcrossed for seven generations.
The samples were prepared by drop casting concentrated NPL solutions onto a zero background silicon substrate.
Negative controls were prepared by omitting the primary antibody, and no staining above the background was detected.
Samples were prepared by drop-casting a concentrated NC solution in chloroform on a low background quartz substrate.
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