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To test the localization of the truncated Ilv enzymes, indirect immunofluorescence microscopy analyses were performed.
(24) The immunofluorescence was conducted to test the localization of ABCG2 protein after treatment with WHI-P154 for 72 h as described previously.
We conclude that a PCNA trimer with all three subunits tagged cannot fulfill all normal PCNA functions, and strains carrying only a tagged version are not suitable to test the localization of PCNA.
To test the localization of NanU, we performed a differential detergent-based cell fractionation and examined the fractions by SDS/PAGE and Western blotting using a BF-NanU antiserum, which we generated from purified His6 NanU from B. fragilis.
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Using O and Oc cells, we tested the localization of the core protein and DDX3 in comparison with IFN-inducing properties (Fig. 3).
At first, we tested the localization of exogenously expressed p35 in recycling endosomes using Alexa 546-transferrin (Tf), which is transported to recycling endosomes when incorporated into cells.
In light of the fact that ethanol dehydration of paraffin-embedded tissue section can lead to tissue shrinkage, we also tested the localization of PADI6, MATER and MSY2 by cryosection analysis, which does not include a dehydration step.
Finally, as alterations of various steps in yolk production are also associated with an egg collapse phenotype [32], [33] we tested the localization of the vitellogenin receptor, Yolkless, in mutant egg chambers in order to assess vitellogenin endocytosis.
Although a transformation system for T. pseudonana has been recently developed [16], we tested the localization of nuPsb28 in the pennate diatom P. tricornutum because transfection is more routine [17], [18].
To determine whether this reliance upon A-type lamins exists in Drosophila, we tested the localization of chromatin and nuclear envelope proteins in the absence of the Drosophila A-type lamin, Lamin C. Previously we isolated two Lamin C null alleles, Lamin CEX296 and Lamin CEX187, which exhibit a broad lethal phase starting at the second instar and ending at the pre-pupal stage [28].
Since the majority of SAS6L-producing eukaryotes do not have a conoid, we tested the localization of a SAS6L ortholog in a distantly related organism.
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