<p><b>Copyright information:</b></p><p>Taken from "Basal shuttle of NF-κB/IκBα in resting T lymphocytes regulates HIV-1 LTR dependent expression"</p><p>http://www.retrovirology.com/content/4/1/56</p><p>Retrovirology 2007;4():56-56.</p><p>Published online 8 Aug 2007</p><p>PMCID:PMC1988826.</p><p></p>�B/IκBα complexes in the nucleus after treatment with LMB or PMA. Ten micrograms of cytosolic and nuclear extracts from CDT cells treated with either PMA or LMB during 4 and 6 hours respectively were analyzed by Western Blot using antibodies against p65/RelA and IκBα. Immunoprecipitation assays were performed using 100 μg of these cytosolic and nuclear extracts, which were incubated with 5 μg of an antibody against p65/RelA conjugated with agarose. IκBα and p65/RelA complexes were characterized by immunoblotting. (b) Contamination with cytosolic proteins during nuclear protein extraction or accumulation of cytosolic proteins in the nucleus after treatment with LMB was assessed by Western Blot using an antibody against both p105 and p50/NF-κB1 proteins. (c) Analysis of NF-κB DNA-binding activity in CDT cells treated with either PMA or LMB. Three micrograms of nuclear extract were incubated with an oligonucleotide containing the double consensus motif κB present in the HIV LTR labeled with [α-P]-dCTP. Protein extracts were obtained from CDT cells after treatment with either LMB or PMA for 6 and 4 hours respectively. (d) Analysis of IκBα pool dependence on protein synthesis. Ten micrograms of nuclear extracts from CDT cells incubated with 20 nM LMB for 4 hours and 10 μg/ml CHX and/or 25 ng/ml PMA for 4 hours,30 min and 2 hours, respectively, were analyzed by Western Blot
