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Blood, Vol. 111, Issue 1, 292-301, January 1, 2008
Down-regulation of the TGF-beta target gene, PTPRK, by the Epstein-Barr virus–encoded EBNA1 contributes to the growth and survival of Hodgkin lymphoma cells
Blood Flavell et al.
111: 292
Supplemental materials for: Flavell et al
Files in this Data Supplement:
- Table S1. EBV-regulated genes in KMH2 (XLS, 627 KB)
- Table S2. EBV-regulated genes in L591 (XLS, 633 KB)
- Figure S1. Q-PCR analysis of PTPRK expression in HL cells (JPG, 36.8 KB)
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(A) Downregulation of PTPRK mRNA expression in KM-H2 cells infected with EBV (KM-H2-EBV) compared with control KM-H2 cells (KM-H2-neo). (B) Downregulation of PTPRK expression in EBV-positive parental L591 cells compared with their EBV-negative counterparts (L591-SD3). (C) EBNA1 expression in EBV-negative cells was alone sufficient to induce the downregulation of PTPRK mRNA expression. Shown here is data for KM-H2 cells. These data provide confirmation of the semi-quantitative PCR analysis shown in Figure 1 of the main paper.
- Figure S2. Q-PCR analysis of PTPRK and Smad2 in HL cells (JPG, 22.8 KB)
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EBV infection (left panel) or EBNA1 expression (right panel) in EBV-negative HL cell lines had no effect on Smad2 mRNA levels. Shown here is data for KM-H2 cells. These findings provide confirmation of the semi-quantitative PCR analysis of Smad2 RNA reported in Figure 2B of the main paper.
- Figure S3. Q-PCR analysis following knockdown of Smad2 in KMH2 neo cells (JPG, 26.0 KB)
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Depletion of Smad2 expression in EBV-negative KMH2 cells following knockdown with siRNA is confirmed by Q-PCR (left panel). This led to the down-regulation of PTPRK expression (right panel). These results confirm the semi-quantitative PCR analysis reported in Figure 4 of the main paper.
- Figure S4. Immunoblotting for EBNA1 (JPG, 11.8 KB)
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Left panel: EBNA1 protein expression is higher in the lymphoblastoid cell line X50-7 than in KM-H2-EBV and EBV-positive L591 cells. Right panel: EBNA1 protein expression in HL cells transfected with EBNA1 is similar to that observed in X50-7. We conclude that EBNA1 is overexpressed in transfected cell lines relative to EBV-infected HL cells. The different size of EBNA1 in the transfected cells reflects the well-described variation in the length of EBNA1 between different EBV isolates.
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