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Blood, Vol. 107, Issue 8, 3321-3329, April 15, 2006
Semaphorin-3A is expressed by tumor cells and alters T-cell signal transduction and function
Blood Catalano et al.
107: 3321
Supplemental materials for: Catalano et al
Files in this Data Supplement:
- Figure S1 (PDF, 52.4 KB) -
T cells were cultured with mitogenic concentrations of soluble anti-CD3 (300 ng/ml) either alone or with anti-CD28 (200 ng/ml). Increasing amounts of human recombinant Sema-3A (Sema3A/Fc) or Sema-6A (Sema6A/Fc) fusion protein were simultaneously added to the culture. After 18 h, cells were pulsed with [3H]-thymidine and T-cell proliferation was measured by thymidine incorporation. Data are the means of triplicate cultures and are representative of three independent experiments.
- Figure S2 (PDF, 91.9 KB) -
Effects of Sema-3A siRNA on mRNA levels of Sema-3A and -3E. Caki-1 and H28 cells were transfected with C-siRNA or Sema-3A siRNA (600 nM). After 48 h, total RNA (50 ng/µl) was isolated and subjected to real-time PCR to detect Sema-3A and Sema-3E transcripts. The mRNA levels of each gene were normalized by the mRNA levels of GAPDH. Reported values are an average of three separate samples of each type analyzed in triplicate; bars, ±SE. *P≤0.001 versus untransfected cells, ANOVA.
- Figure S3 (PDF, 92.8 KB) -
Jurkat T cells were infected with an empty vector or oncogenic ras (Ha-RasV12) as described in “Materials and methods.” After 4 days, (A) infected-Jurkat cells were stimulated with Sema3A/Fc or Sema6A/Fc fusion protein (100 ng/ml) for the indicated times. Total cell lysates were prepared and immunoblotted with a specific antibody against phospho-ERK1/1 (anti-p-ERK1/2) or with anti-ERK1 antibody as a loading control. Data are representative of two independent experiments with comparable results. (B) Infected Jurkat T cells were cultured in the absence or presence of Sema3A/Fc (100 ng/ml) for additional 18 h, and then cell proliferation was measured by thymidine incorporation. Data are the means of triplicate and are representative of three independent experiments.
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