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. Author manuscript; available in PMC: 2010 Jun 1.
Published in final edited form as: Cancer Res. 2009 May 19;69(11):4589–4597. doi: 10.1158/0008-5472.CAN-08-4117

Figure 1. The c-Rel and v-Rel subunits of NFκB associate with Pin1.

Figure 1

(A) Sequence alignment of the vertebrate Rel/NF-κB family proteins, highlighting conservation of N-terminal sequences flanking the Thr254-Pro Pin1 recognition motif in RelA, whereas sequences flanking its C-terminal end are more divergent. (B) Immunoblots showing endogenous expression of Pin1 in 293T cells, v-Rel-transformed CSC and in the primary mediastinal B cell lymphoma cell line Karpas 1106. The blot was probed with anti-Pin1 and reprobed for actin. (C) Left panel. Pull-down of v-Rel or hc-Rel in extracts from transiently transfected 293T cells with GST-Pin1 or GST as control, followed by immunoblotting with anti-Rel. Where indicated, cells were treated with hTNFα prior to harvest (lanes 4–6). Right panel. Pull-down of v-Rel, hc-Rel or hRelA transfected in 293T cells with GST-Pin1, followed by immunoblotting with anti-Rel. Where indicated, cells were treated with hTNFα. Input (1/10 of lysate; 1–2, 5–6 and 9–10). The blot was reprobed for actin and Pin1. (D) Left panel. Pull-down of endogenous v-Rel from v-Rel-transformed CSC (lanes 1–3) or hc-Rel from human lymphoma-derived Karpas 1106 cells (lanes 4–6) with GST-Pin1 or GST, followed by immunoblotting with anti-Rel (v-Rel: #1691 and hc-Rel: #265). Right panel. Co-immunoprecipitation of endogenous hc-Rel with Pin1 in extracts from human lymphoma cell lines using anti-Pin1, followed by immunoblotting with anti-hc-Rel. Input (1/10 of lysate). The membrane was stained with Ponceau S (bottom panel).