Author: Qiao, Hui; Pelletier, Sandra L.; Hoffman, Lucas; Hacker, Jill; Armstrong, R. Todd; White, Judith M.
Title: Specific Single or Double Proline Substitutions in the “Spring-loaded” Coiled-Coil Region of the Influenza Hemagglutinin Impair or Abolish Membrane Fusion Activity Document date: 1998_6_15
ID: 78fjem8s_12
Snippet: As indicated, cells expressing wt-or mutant HAs were washed twice with RPMI or PBS and incubated for 6 min at room temperature (RT) with RPMI or PBS containing either TPCK-trypsin (5 g/ml) or, as a negative control, TLCK-chymotrypsin (5 g/ml) (Sigma Chemical Co., St. Louis, MO). Trypsin-treated cells were then incubated for 10 min with RPMI or PBS containing soybean trypsin inhibitor (50 g/ml). Higher concentrations of trypsin (10 g/ml) were used.....
Document: As indicated, cells expressing wt-or mutant HAs were washed twice with RPMI or PBS and incubated for 6 min at room temperature (RT) with RPMI or PBS containing either TPCK-trypsin (5 g/ml) or, as a negative control, TLCK-chymotrypsin (5 g/ml) (Sigma Chemical Co., St. Louis, MO). Trypsin-treated cells were then incubated for 10 min with RPMI or PBS containing soybean trypsin inhibitor (50 g/ml). Higher concentrations of trypsin (10 g/ml) were used where noted to increase the amount of cleaved HA at the cell surface.
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