Table 3.

Inhibition of pp59Lyn by neutralizing antibody in adipocytesa

Antibody dilutionPhosphorylation (arbitrary units)
BasalPIG 41Insulin
None8,513 ± 1,11294,672 ± 12,73232,462 ± 4,244
Anti-rat IgG (1:25)8,034 ± 1,03491,207 ± 13,24535,785 ± 4,890
Anti-pp59Lyn
 1:2,0008,744 ± 1,05585,992 ± 11,30839,833 ± 4,067
 1:5007,231 ± 86550,315 ± 7,85523,121 ± 3,133
 1:1006,023 ± 76237,312 ± 2,98819,452 ± 2,629
 1:505,149 ± 44127,057 ± 3,56210,038 ± 1,759
 1:254,540 ± 33123,560 ± 2,2348,207 ± 1,552
 1:104,267 ± 25520,135 ± 2,4337,064 ± 970
  • a Isolated rat adipocytes were electroporated with neutralizing anti-pp59Lyn or anti-rat IgG antibody at various dilutions and then incubated (15 min, 37°C) in the absence or presence of PIG 41 (3 μM) or human insulin (10 nM). pp59Lyn was immunoprecipitated with monoclonal anti-pp59Lyn antibody from defatted cell lysates and then immunoblotted with antiphosphotyrosine antibody. Autophosphorylated kinase was quantitatively evaluated by phosphorimaging (arbitrary units). The experiment was repeated twice (mean ± standard deviation).