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. 1999 Jul 19;190(2):281–292. doi: 10.1084/jem.190.2.281

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© 1999 The Rockefeller University Press

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Expression and binding of endogenous SAF. (A) Western blot analysis with the SAF antisera (top) and the control anti–β-actin antibody (bottom). Western blot analysis on whole cell extracts from 293T cells (lane 1): 293T cells transfected with 5, 10, and 20 μg of the CMV–SAF expression construct (lanes 2–4); the CD4⁺CD8⁺ DP AKR1G1 thymoma (lane 6); the CD8⁺ SP Tc L3 cell clone (lane 7); the CD4⁺ SP Th D10 cell clone (lanes 8 and 9); the P3X63 plasmacytoma (lane 10); the 103 pre-B cell lymphoma (lane 11); the WEHI-3B macrophage/monocyte (lane 12); and the 3T3 fibroblast (lane 13). Molecular weight standards were loaded in lane 5. (B) Antibody ablation EMSA analyses with the SAF antisera and (left) CD4⁺ SP Th D10 or (right) CD8⁺ SP Tc L3 extracts. Lanes containing S3 probe only, S3 probe with extract, and S3 probe with extract and antibody are indicated above each lane. ‘Pre’ indicates preimmune sera.

Expression and binding of endogenous SAF. (A) Western blot analysis with the SAF antisera (top) and the control anti–β-actin antibody (bottom). Western blot analysis on whole cell extracts from 293T cells (lane 1): 293T cells transfected with 5, 10, and 20 μg of the CMV–SAF expression construct (lanes 2–4); the CD4⁺CD8⁺ DP AKR1G1 thymoma (lane 6); the CD8⁺ SP Tc L3 cell clone (lane 7); the CD4⁺ SP Th D10 cell clone (lanes 8 and 9); the P3X63 plasmacytoma (lane 10); the 103 pre-B cell lymphoma (lane 11); the WEHI-3B macrophage/monocyte (lane 12); and the 3T3 fibroblast (lane 13). Molecular weight standards were loaded in lane 5. (B) Antibody ablation EMSA analyses with the SAF antisera and (left) CD4⁺ SP Th D10 or (right) CD8⁺ SP Tc L3 extracts. Lanes containing S3 probe only, S3 probe with extract, and S3 probe with extract and antibody are indicated above each lane. ‘Pre’ indicates preimmune sera.