Journal: Oncogene

Article Title: A chicken c-Rel-estrogen receptor chimeric protein shows conditional nuclear localization, DNA binding, transformation and transcriptional activation.

doi: 10.1038/sj.onc.1201860

Figure Lengend Snippet: Figure 3 Subcellular localization of Rel proteins in control untransfected CEF (a), c-Rel CEF (b), Dc-Rel CEF (c), c-RelER CEF (d-g), and Dc-RelER CEF (h and i). c-RelER CEF were incubated with estrogen, OHT, or ICI (e, f, g, respectively), or without drug (d). Dc-RelER CEF were incubated with estrogen (i) or without (h). Fixed and permeabilized cells were incubated with the SB66 anti-Rel antibody, followed by incubation with ¯uorescein-conjugated anti-rabbit immunoglobulin

Article Snippet: CEF were preincubated in methionine- and cysteine-free medium and labeled with 35S-trans Label (Amersham) for 2 h. Cells were lysed in 0.5% NP40, 0.5% deoxycholate, 20 mM Tris-HCl, pH 8.0, 50 mM NaCl and subjected to immunoprecipitation analysis using the Rel-speci®c SB66 antibody, and HC-20, a rabbit polyclonal antibody raised against the hormone-binding domain of the human estrogen receptor (Santa Cruz Biotechnology), as previously described (Morrison et al., 1989).

Techniques: Control, Incubation

Journal: Oncogene

Article Title: A chicken c-Rel-estrogen receptor chimeric protein shows conditional nuclear localization, DNA binding, transformation and transcriptional activation.

doi: 10.1038/sj.onc.1201860

Figure Lengend Snippet: Figure 4 c-RelER forms speci®c complexes with cellular proteins. Protein extracts prepared from 35S-methionine-labeled c-Rel CEF (lanes 1, 7 and 13), control untransfected CEF (lanes 6, 12 and 18), and c-RelER CEF grown in the presence of estrogen (lanes 3, 9 and 15), ICI (4, 10 and 16), OHT (lanes 5, 11 and 17), or mock treated cells (lanes 2, 8 and 14), were immunoprecipitated with the HC-20 anti-estrogen receptor antibody (lanes 1 ± 6), SB66 anti-Rel antibody (lanes 7 ± 11), or preimmune sera (lanes 13 ± 18). The positions of the proteins which coprecipitate with c-RelER are indicated on the left

Article Snippet: CEF were preincubated in methionine- and cysteine-free medium and labeled with 35S-trans Label (Amersham) for 2 h. Cells were lysed in 0.5% NP40, 0.5% deoxycholate, 20 mM Tris-HCl, pH 8.0, 50 mM NaCl and subjected to immunoprecipitation analysis using the Rel-speci®c SB66 antibody, and HC-20, a rabbit polyclonal antibody raised against the hormone-binding domain of the human estrogen receptor (Santa Cruz Biotechnology), as previously described (Morrison et al., 1989).

Techniques: Labeling, Control, Immunoprecipitation