Materials and Reagents

1. Cell labeling dyes
   1. Vybrant CM-DiI (Life Technologies, Invitrogen^TM, catalog number: V-22888 )
      
   2. VybrantDiD (Life Technologies, Invitrogen^TM, catalog number: V-22887 )
      
   3. CellTrace CFSE (Life Technologies, Invitrogen^TM, catalog number: C-34554 ) (either CM-DiI or CFSE can be used based on the preferred wave length either FL-1 for CFSE or FL-2 for the CM-DiI)
      
2. Other materials
   1. Phosphate buffered saline (PBS)
      
   2. 1x Dulbecco’s Modified Eagle Medium (DMEM) (Invitrogen,Life Technologies, Invitrogen^TM, catalog number: 10313-039 )
      
   3. Fetal bovine serum (Life Technologies, Invitrogen^TM, catalog number: 11091-148 )
      
   4. 40 micron cell strainer (BD Biosciences, Falcon^®, catalog number: 352340 )
      
   5. Mice of interest
      
   6. NaN₃ (Sodium Azide)
   7. Erythrocyte lysis buffer (ELB) (see Recipes)
      
   8. FACS buffer (see Recipes)
      
3. Antibodies
   To demonstrate that the observed effect is specific we recommend to either blocking the relevant receptors, or by depleting NK cells, by using anti-NK1.1 (in case the receptor is present), or by using anti-asialo GM1.
   1. Anti NK1.1 (eBioscience, catalog number: 14-5941-82 )
      
   2. Anti-Asialo-GM1 (eBioscience, catalog number: 16-6507-39 )
      
   3. Anti-Mouse CD314 (NKG2D) (eBioscience, catalog number: 14-5873-82 )
      

Equipment

1. BD LSR II flow cytometer

Procedure