Proprotein convertase furin is preferentially expressed in T helper 1 cells and regulates interferon gamma
Blood Pesu et al. 108: 983

Supplemental materials for: Pesu et al, Vol 108, Issue 3, 983-985

Files in this Data Supplement:

• Figure S1. Inactive furin mutant does not upregulate IFN-γ (PDF, 11.5 KB) -
  Furin-deficient LoVo cells were transfected with 0.3 µg human IFN- cDNA and/or 1.7 µg wild-type or D153A mutant furin cDNA. pcDNA3 vector was used to equalize DNA concentrations. Seventy-two hours after transfection, secreted IFN- protein was measured by ELISA. The experiment was performed twice with similar results.
  
  
• Figure S2. Furin inhibition blocks intracellular IFN-γ expression (PDF, 40.3 KB) -
  Human CD3⁺ T cells were prepared as described in “Study design” and rested overnight in serum-free media. Cells were stimulated for 24 hours with 100 ng/mL IL-12 in the presence of ∝₁-antitrypsin or ∝₁-PDX (7 µM). Golgistop was added to cell culture for the last 4 hours of stimulation. Cells were surface stained with anti—CD4-APC and fixed in 4% formyl saline, followed by intracellular staining with anti—IFN--FITC in 0.1% saponin permeablization buffer. CD4 and IFN- expressions were analyzed with FACS Calibur (Becton Dickinson). The experiment was performed twice with similar results.
  
  
• Figure S3. Furin inhibition interferes with IL-12+18 induced IFN-γ production (PDF, 10.8 KB) -
  Human T cells were prepared as in Figure 1 and stimulated with IL-12 and IL-18 (50 + 50 ng/mL for 65 hours) in the presence of ∝₁-antitrypsin or ∝₁-PDX (20 µM). IFN- production was analyzed with ELISA. The experiment was performed twice with similar results.