Materials and Reagents

1. 7AAD (7-amino-actinomycin D) (BD Pharmingen, catalog number: 51-68981E )
   
2. Bovine serum albumin (BSA) (PAA Laboratories GmbH, catalog number: K41-001 )
   
3. DMEM (Life Technologies, Gibco^®, catalog number: 41966-029 )
   
4. Superior FBS (fetal bovine serum, not heat-inactivated) (Biochrom, catalog number: S0615 )
   
5. Mx/RAGE7 cells (virus-transformed adherent cell line with a temperature-inducible promotor; must be cultured at 32 °C; cells express the Mx transgene and a promotorless eGFP gene which is expressed due to type I IFN stimulation ) (Bollati-Fogolin and Muller, 2005)
   
6. PBS (Life Technologies, Gibco^®, catalog number: 14190-136 )
   
7. Penicillin/streptomycin (PAA Laboratories GmbH, catalog number: P11-010 )
   
8. Propidium iodide (eBioscience, catalog number: 00-6990-50 )
   
9. Murine IFN-α (PBL, catalog number: 12100-1 )
   
10. Sodium azide (Applichem, catalog number: A1430.0010 )
    
11. Sodium pyruvate (Life Technologies, Gibco^®, catalog number: 11360-039 )
    
12. Trypsin EDTA (PAA Laboratories GmbH, catalog number: L11-004 )
    
13. β-mercaptoethanol (Life Technologies, Gibco^®, catalog number: 31350-010 )
    
14. Media for Mx/RAGE7 cells (see Recipes)
    
15. FACS buffer (see Recipes)
    

Equipment

1. 96-well flat bottom plate (Falcon BD Labware, catalog number: 3072 )
   
2. 1.5 ml microfuge tubes
   
3. FACS tubes (BD Biosciences, Falcon^®, catalog number: 352054 )
   
4. Flow cytometer (e.g. BD LSR II)
   
5. Incubator (37 °C; 5% CO₂)
   
6. Incubator (32 °C; 5% CO₂)
   

Procedure