Table 1

Summary of the phenotypes of the p60ΔCD mutants (16).

MAB225*Clone 4.12TNF-α bindingSelfassociation§Dominant interference|
p60ΔCD111++
p6055-211 0.870.020.01NT
K19E1.051.071.1NT+
KY19/20AA0.590.140.03
Q24A1.060.971.13++
K32A0.360.010.01
DT49/50AA1.160.961.13NT+
E57A1.680.040.02++
T61A1.421.281.35NT+
N66F0.670.040.01++
R77A1.060.991.24NT+
W108T1.321.111.25NT+
L112E1.311.270.85NT+
  • * Staining of p60-specific monoclonal antibody clone MAB225 (R&D Systems). The values were normalized against the staining of the HA epitope tag by dividing the percentage of MAB225-positive cells by the percentage of HA-positive cells.

  • Staining with p60-specifc monoclonal antibody clone 4.12 (Zymed) was normalized against HA staining.

  • TNF-α binding was determined with a biotinylated form of TNF-α and normalized against HA staining (26).

  • § Self-association was determined by immunoprecipitation assays in 293T transient transfections as described (12). NT, not tested.

  • | Dominant interference was determined as described (27). Dominant inhibition by the p60ΔCD-HA mutants was at least 50% of p60ΔCD wild type (+). p6055-211, KY19/20AA, and K32A did not confer any protection (<5%) against TNF-induced death (−) in all experiments. The antibodies and TNF-α binding to p60ΔCD are arbitrarily set at 1. Results are representative of three independent experiments.