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path4198-sup-0001-Appendix S1.docWord document44KAppendix S1. Supplementary materials and methods
path4198-sup-0002-Figure S1.tifTIFF image617KPhenotype of enriched CD10+ germinal centre B cells. Flow-cytometric analysis of tonsillar mononuclear cells after enrichment for CD10. (A) Cells were stained for CD10 or CD19 together with CD24. (B) Cells were stained for CD10, CXCR4 and CD83. (C) Cells were stained for CXCR4 and CD77
path4198-sup-0003-Figure S2.tifTIFF image2366KExpression of LMP2A in primary human germinal centre B cells. (A) Flow-cytometric analysis of CD10+ tonsillar B cells transfected with pSG5 (left) or LMP2A (right), together with pMACSLNGFR and stained for CD10 and NGFR prior to sorting (B) Detection of LMP2A and β-actin in transfected CD10+ tonsillar B cells by immunoblot analysis prior to sorting
path4198-sup-0004-Figure S3.pdfPDF document262KLMP2A and BCR activation regulate a common subset of target genes in primary human GC and BL2 cells. LMP2A target genes were compared with those genes shown to be differentially expressed in BCR crosslinked GC cells compared to unstimulated cells. (A) 323 genes were up-regulated by LMP2A in GC B cells and 1536 in stimulated GC B cells. Genes up-regulated following BCR activation were substantially enriched for those up-regulated by LMP2A [odds ratio (OR) = 16, p < 0.0001]. (B) 463 genes were down-regulated by LMP2A in GC B cells and 1572 in stimulated GC B cells. Genes down-regulated by BCR activation were enriched for those down-regulated by LMP2A (OR = 14.7, p < 0.0001). (C) qRT–PCR of the relative quantity of CCL3, CCL4, XBP1 and AID in LMP2A-expressing and control-transfected CD10+ GC B cells in two different tonsil samples (left panels), or following BCR crosslinking or CD40 stimulation in GC B cells (right panels). (D) Expression of these genes in LMP2A transfected BL2 cells. (E) Real-time RT–PCR of the relative quantity of STAT1, IFITM1, IFI6, HERC5 and CD80 in LMP2A-expressing and control-transfected CD10+ GC B cells in two different tonsil samples
path4198-sup-0005-Figure S4.tifTIFF image160KLMP2A expression does not induces activation of the transcription factor Elk-1 in L1236 HL cells. L1236 cells were transfected with pSG5, pSG5–LMP2A or pFC–MEKK1 together with pFA–Elk1 and pFR–Luc. Relative luciferase activity was determined 24 h post-transfection. The results are the mean of three independent experiments (± SD)
path4198-sup-0006-Figure S5.tifTIFF image826KOverlap between transcriptional targets of LMP1, LMP2A, BCR crosslink and primary HRS cells compared to centrocytes. LMP2A and BCR target genes were compared with those genes shown to be differentially expressed in HRS cells compared to centrocytes and LMP1-expressing GC B cells. (A) 258 genes were up-regulated by LMP2A and BCR crosslink in GC B cells and 172 by LMP1 and in HRS cells compared to centrocytes. (B) 235 genes were down-regulated by LMP2A and BCR crosslink in GC B cells and 537 by LMP1 and in HRS cells compared to centrocytes
path4198-sup-0007-Table S1.xlsxExcel 2007 spreadsheet68KGenes significantly up- or down- regulated by LMP2A in GC B cells with fold change > 1.3 and p value < 0.01.
path4198-sup-0008-Table S2.xlsxExcel 2007 spreadsheet236KGenes significantly up- or down- regulated by BCR in GC B cells with fold change > 1.3 and p value < 0.01.
path4198-sup-0009-Table S3.xlsxExcel 2007 spreadsheet25KList of genes that are concordantly regulated by LMP2A and BCR in GC B cells.

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