Materials and Reagents

1. Human foreskin fibroblasts (HFFs) cells culture (ATTC, catalog number: SCRC-1042 )
   
2. Dulbecco's modified Eagle medium (DMEM) (Life Technologies, catalog number: 41966-029/052 )
   
3. Fetal bovine serum (FBS) (Eurobio^®, catalog number: CVFSVF0001 )
   
4. Penicillin/streptomycin (PAN Biotech GmbH, catalog number: P0607-100 )
   
5. L-Glutamine (200 mM) (Life Technologies, catalog number: 25030-024 )
   
6. Dulbecco's phosphate-buffered saline (DPBS) (Life Technologies, catalog number: 14190-094/069 )
   
7. Proteinase K (molecular biology grade) (Bio-Rad Laboratories, catalog number: P8107S , Lot: 0051310)
   
8. TRIS (Euromedex, catalog number: 26-128-3094-B )
   
9. EDTA (Euromedex, catalog number: E013 )
   
10. SDS (Euromedex, catalog number: EU0660 )
    
11. NaCl (Euromedex, catalog number: 1112-A )
    
12. HCl (37% ACS reagent) (Sigma-Aldrich, catalog number: 258148-2.5ML )
    
13. Phenylmethylsulfonylfluoride (PMSF) (Euromedex, catalog number: 1111-C )
    
14. Hoescht 33258 (Life Technologies, Molecular Probes^®, catalog number: H-3569 )
    
15. Formaldehyde methanol free (Polysciences, catalog number: 0 4018 )
    
16. Dolichos biflorus lectin coupled to fluorescein isothiocyanate (FITC) (Clinisciences, catalog number: FL-1031 )
    
17. Complete DMEM medium (see Recipes)
    
18. Phenylmethylsulfonylfluoride (PMSF) solution (see Recipes)
    
19. Lysis buffer (see Recipes)
    

Equipment

1. 37 °C/5% CO₂ cell culture incubator
   
2. 6-well plates
   
3. Scissors and forceps
   
4. Glass homogenizers (Potter-Elvehjem PTFE, 15 ml) (Dutscher, catalog numbers: 057009 and 057021 )
   
5. 15 ml polystyrene tubes
   
6. Epifluorescence microscope
   

Procedure