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Examination of changes in structural features of the human telomere G-quadruplex resulting from UP1 binding were performed using CD spectrophotometry.
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Visualisation of binding was performed using ImageQuantTL v8.1.0.0 (GE Healthcare, Buckinghamshire, UK).
Direct synthesis of [99mTc]TMCE from dimethyl iminodiacetate, which contains both the asymmetric urea and succinimidyl moiety important for PSMA binding, was performed using our microwave-assisted one-pot procedure.
High affinity binding was performed using I-APE as described above.
Detection of antibody binding was performed using the Envision HRP kit K4061 (Dako).
24 hr after splitting, binding was performed using AP-tagged ligands (AP-VEGF A, AP-SEMA3A, AP-VEGF B, AP-PlGF).
In silico analysis of transcription factor binding was performed using the JASPAR (http://jaspar.genereg.net) open-access database of transcription factor binding profiles (41).
NF-κB DNA binding was performed using a duplex oligonucleotide probe containing the consensus NF-κB binding sequence (Han et al, 2009a; Ma et al, 2004).
Filter binding was performed using nitrocellulose filters followed by three washes with 5% (w/v) chilled TCA, and radioactivity was determined.
AP-RR36 in situ binding was performed using AP fluorescent substrates (Fig. 1A,B) or followed by double immunofluorescence labeling of AP and several NPCs markers (Fig. 1A,C N).
Blocking a non-specific binding was performed using either 3% fatty acid-free bovine serum albumin or 5% non-fat dry milk for 3 h.
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