36 



dispersed by pasteur pipetting with a silicone treated 

 pipette. A single plate containing 18 individual wells was 

 processed at a time with the entire procedure requiring less 

 than 30 minutes. 



Inhibition of H-7 and H-7Wcr cell growth by Con A, 

 ADGG or ADGG-Con A conjugates was measured by exposure of 



established cell monolayers to the protein for 15 minutes. 



4 2 



Cells were plated at a density of 1 x 10 cells/cm and 



allowed to establish growth for 12 hours. The media was 

 carefully aspirated and the cells gently rinsed two times 

 with 0.15M PBS, pH 7.4 at room temperature. Dilutions of 

 each inhibitor were added in 1.0ml of PBS to triplicate 

 wells for each concentration used. After 15 minutes of 

 exposure, the protein solution was aspirated, the cells 

 rinsed twice with PBS and 1.0ml fresh media added to the 

 well. After 48 hours of growth, the cells were processed 

 for counting as described above. 



A- 9 and LAN-2 cells were treated as described for 

 CHO H-7 and H-7Wcr cells for determination of their sensi- 

 tivity to free or conjugated cx-amanitin. 



Inhibition of H-Thymidine Incorporation 



3 3 



Incorporation of [methyl- H] -thymidine ( H-TdR) was 



measured for AV3, CHO M-7 and EL4 cells as an additional 



method for assessing inhibition of cellular functions by 



ADH-BSA conjugates. A procedure similar to that of Ball, 



et al. (1973) was used. Labeled precursor (Schwartz-Mann, 



5mCi/mmole) was added as a lOOyl addition to give a final 



