NFκB activity, function, and target-gene signatures in primary mediastinal large B-cell lymphoma and diffuse large B-cell lymphoma subtypes by Friedrich.

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NFκB activity, function, and target-gene signatures in primary mediastinal large B-cell lymphoma and diffuse large B-cell lymphoma subtypes by Friedrich Feuerhake, Jeffery L. Kutok, Stefano Monti, Wen Chen, Ann S. LaCasce, Giorgio Cattoretti, Paul Kurtin, Geraldine S. Pinkus, Laurence de Leval, Nancy L. Harris, Kerry J. Savage, Donna Neuberg, Thomas M. Habermann, Riccardo Dalla-Favera, Todd R. Golub, Jon C. Aster, and Margaret A. Shipp Blood Volume 106(4):1392-1399 August 15, 2005 ©2005 by American Society of Hematology

Subcellular localization of c-REL in MLBCL Subcellular localization of c-REL in MLBCL. (A) Prominent intense c-REL nuclear staining in a representative primary MLBCL. (B) Variable c-REL staining intensity and subcellular localization in a DLBCL. Original magnification, × 400. Subcellular localization of c-REL in MLBCL. (A) Prominent intense c-REL nuclear staining in a representative primary MLBCL. (B) Variable c-REL staining intensity and subcellular localization in a DLBCL. Original magnification, × 400. Friedrich Feuerhake et al. Blood 2005;106:1392-1399 ©2005 by American Society of Hematology

NFκB activity and inhibition in an MLBCL cell line. NFκB activity and inhibition in an MLBCL cell line. (A) NFκB DNA-binding activity of the MLBCL cell line, Karpas 1106; 2 DLBCL cell lines, OCI LY10 and DHL6; and a Hodgkin lymphoma cell line, KM-H2. NFκB DNA-binding activity was measured using a colorimetric assay. Measurements were performed in triplicates. (B) NFκB activity and apoptosis in MLBCLs expressing an IκBα superrepressor. (Left) NFκB DNA-binding activity in MLBCL cells transduced with MSCV-eGFP (vector) alone or MSCV-eGFP-SR-IκBα. Measurements were performed in triplicate. NFκB activity was significantly lower in SR-IκBα cells than in vector-only cells (P < .001, one-sided Student t test). (Right) The apoptotic fraction (percentage annexin V+ GFP+) of cells transduced with MSCV-eGFP-SR-IκBα is much higher than that of cells transduced with vector alone. (C) Apoptosis in MLBCL cells expressing an IκB superrepressor. GFP-positive cells expressing vector alone or MSCV-eGFP-SR-IκBα were analyzed for expression of annexin V (Alexa 568). GFP, x-axis; annexin V, y-axis. (D) Proliferation of MLBCL cells expressing an IκBα superrepressor. The proliferation (MTS absorbance) of parental and GFP+ vector-only- and MSCV-eGFP-SR-IκBα-transduced MLBCL cells was measured at 0, 24, 48, and 72 hours and assessed with an ANOVA model that included the type of treatment and time of measurement. The difference between SR-IκBα-transduced MLBCL cells and either vector-only or parental cells was significant (P < .001 in each case), whereas there was no difference between empty vector and parental cells (P = NS). The measurements were performed in triplicate. All experiments (A-D) were performed 3 to 4 times with comparable results; representative experiments are shown. Error bars indicate the standard deviation within triplicate experiments. Friedrich Feuerhake et al. Blood 2005;106:1392-1399 ©2005 by American Society of Hematology

Differential expression of NFκB target genes in large B-cell lymphoma subtypes. Differential expression of NFκB target genes in large B-cell lymphoma subtypes. NFκB target genes that met the 99th percentile of the permutation distribution and exhibited a 30% or more difference in median expression values were considered to be differentially expressed. Differentially expressed NFκB target genes in (A) primary MLBCL versus DLBCL; (B) ABC-like DLBCLs versus GCB-like and other DLBCLs; and (C) HR DLBCLs versus OxPhos and BCR/proliferation DLBCLs. (D) Comparison of the NFκB target gene signatures from primary MLBCL, HR DLBCLs, and ABC-like DLBCLs. Friedrich Feuerhake et al. Blood 2005;106:1392-1399 ©2005 by American Society of Hematology

Differential expression of NFκB target genes in an independent series of primary MLBCLs and DLBCLs. The independent data set includes all primary MLBCLs (38 tumors) and DLBCLs (26 tumors: 13 GC-type and 13 ABC-like) that were made available at the NIH Lymph... Differential expression of NFκB target genes in an independent series of primary MLBCLs and DLBCLs. The independent data set includes all primary MLBCLs (38 tumors) and DLBCLs (26 tumors: 13 GC-type and 13 ABC-like) that were made available at the NIH Lymphoma/Leukemia Molecular Profiling Project website.27 Differentially expressed NFκB target genes were defined as in Figure 3. (A) MLBCLs versus DLBCLs. This independent series of primary MLBCLs had significantly higher expression of 79% (26/33) of the NFκB target genes that were identified in our series and represented on both platforms; and (B) GC-type versus ABC-like DLBCLs. Friedrich Feuerhake et al. Blood 2005;106:1392-1399 ©2005 by American Society of Hematology

cREL amplification and transcript abundance in large B-cell lymphoma subtypes. cREL amplification and transcript abundance in large B-cell lymphoma subtypes. (A) c-REL transcript abundance in primary DLBCLs with an amplified cREL locus (positive, 15/127 examined tumors) or no cREL amplification (negative, 112/127 examined tumors). (B) c-REL transcript abundance and cREL amplification in DLBCLs arranged by cell of origin (GCB, ABC, and other) and primary MLBCL. Error bars indicate the standard deviation within triplicate experiments. Friedrich Feuerhake et al. Blood 2005;106:1392-1399 ©2005 by American Society of Hematology