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Blood, Vol. 108, Issue 13, 4178-4186, December 15, 2006
Role of phosphatidylinositol 3'-kinase/AKT pathway in diffuse large B-cell lymphoma survival
Blood Uddin et al.
108: 4178
Supplemental materials for: Uddin et al, Vol 108, Issue 13, 4178-4186
Files in this Data Supplement:
- Table S1. Cox regression analysis for overall survival of patients with DLBCL (PDF, 14.4 KB) -
Multivariate analysis with IPI and p-AKT as predictors of survival was done using Cox proportional hazards regression models. In the univariate Cox regression analysis, the relative risk for death was 1.9 for high p-AKT expression (95% CI, 0.7-5.5; P = .2) and 5.3 for the IPI (95% CI, 1.9-18.7; P = .001). In the multivariate Cox regression analysis that included p-AKT and the IPI, the relative risk for death was 1.4 for high AKT expression (95% CI, 0.5-4.2; P = .451) and 5.0 for the IPI (95% CI, 1.7-17.8; P = .002).
- Figure S1. Cell-cycle analysis of DLBCL cell lines (JPG, 90.5 KB)
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SUDHL4 and SUDHL5 cell lines were treated with 25 µM LY294002 for indicated time periods and cell-cycle fractions were analyzed by flow cytometry.
- Figure S2. Wortmannin and direct AKT inhibitor induced cell death in DLBCL cell lines (JPG, 75.1 KB)
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LY294002-sensitive SUDHL4 and LY294002-resistant SUDHL8 cell lines were treated with indicated doses of wortmannin and direct AKT inhibitor for 24 hours and cell death was analyzed by (A) FITC-conjugated annexin V/PI dual staining and (B) trypan blue exclusion dye.
- Figure S3. LY294002-induced down-regulation of XIAP expression (JPG, 42.8 KB)
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SUDHL4, SUDHL8, and SUDHL10 cells were treated with and without 25 µM LY294002 for 24 hours. Cells were lysed and equal amount of proteins were separated on SDS-PAGE, transferred to PVDF membrane, and immunoblotted with antibodies against XIAP and actin as indicated.
- Figure S4. Detection and expression of p-AKT (JPG, 64.6 KB)
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(A) Detection of constitutively active AKT in DLBCL cell lines. Serum starvation experiments were performed on all DLBCL cell lines. Cells were grown in serum (+) and serum (–) media for 24 hours. Cells were lysed, immunoblotted, and probed with anti-p-AKT and actin antibodies as indicated. (B) Constitutive expression of p-AKT in DLBCL cell lines by immunohistochemistry. SUDHL4, SUDHL10, SUDHL8, and OCI-LY19 cell line blocks were stained for IHC with p-AKT antibody (original magnification, ×40).
- Figure S5. IPI risk groups and p-AKT expression (JPG, 133 KB)
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IPI risk groups showing proportion of DLBCL cases with p-AKT overexpression (low risk 39.13%, intermediate risk 56%, and high risk 75%, P = .079).
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