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. 2015 Mar 16;11(3):e1004721. doi: 10.1371/journal.ppat.1004721

Fig 5. Tax requires CADM1 for NF-κB activation.

Fig 5

(A) Lentiviral Tax was transduced in Jurkat T-cells stably expressing control scrambled shRNA or CADM1 shRNA. After 48 h, lysates were subjected to immunoblotting with anti-IκBα, anti-phospho-IκBα, anti-CADM1, anti-Tax, and anti-β-actin antibodies. (B) Primary Cadm1 +/+ and Cadm1 −/− MEFs were transduced with Tax-expressing lentiviruses. After 48 h, lysates were subjected to immunoblotting with anti-IκBα, anti-phospho-IκBα, anti-CADM1, anti-Tax, and anti-β-actin antibodies. (C) Nuclear extracts from lentiviral expressing Tax in primary Cadm1 +/+ and Cadm1 −/− MEFs were used for NF-κB and Oct-1 EMSA, and cytoplasmic extract were subjected to immunoblotting with anti-Tax, anti-CADM1, and anti-β-actin antibodies. (D) Lysates from HTLV-1 transformed C8166, MT-2, and MT-4 cells stably expressing control scrambled shRNA or CADM1 shRNA were subjected to immunoblotting with anti-IκBα, anti-phospho-IκBα, anti-CADM1, anti-Tax, and anti-β-actin antibodies. (E) Nuclear extracts from HTLV-1 transformed C8166, MT-2, and MT-4 cells stably expressing control scrambled shRNA or CADM1 shRNA were used for NF-κB and Oct-1 EMSA, and cytoplasmic extracts were subjected to immunoblotting with anti-CADM1 and anti-β-actin antibodies. (F) Primary Cadm1 +/+ and Cadm1 −/− MEFs were transduced with Tax-expressing lentiviruses as described for panel B. After 48 hours, RNA was prepared and subjected to RT-PCR for A20, IL-6, Bfl-1, Tax, and GAPDH expression.