Figure 7.

Recovery of PAR₂-mediated signaling. A, experimental design. HEK293 cells were first challenged with trypsin (10 nm), CS (100 nm), NE (100 nm), or vehicle for 10 min. Cells were then washed, recovered for 110 min, and then challenged with trypsin (10 nm) at 120 min after the first challenge with proteases. B–D, measurement of [Ca²⁺]_i in HEK293 cells. B, effects of PKD inhibitor (CRT0066101, CRT, 100 nm) or Gβγ inhibitor (gallein, 10 μm) on responses to a single challenge with trypsin. C and D, responses to a first challenge with trypsin, CS, or vehicle (C) or with trypsin, NE, or vehicle (D) and a second challenge with trypsin after 120 min. n = 4 (B) or 5 (C and D) experimental replicates from triplicate observations. Error bars, S.E.