Figure 1.

Protease-induced activation of PKD. A, localization of FRET PKD biosensors Cyto-DKAR and Golgi-DKAR with TGN58K in HEK293 cells. B–E, time course of trypsin-induced (B, 10 nm), CS-induced (C, 100 nm), NE-induced (D, 100 nm), and 2-furoyl-LIGRLO-NH₂–induced (E, 10 μm) PKD activity in HEK293 cells expressing FRET biosensors for PKD in the cytosol (Cyto-DKAR) and Golgi apparatus (Golgi-DKAR). DKAR-T/A is a PKD control sensor. Agonists were added at the arrow. F–H, AUC for Cyto-DKAR and DKAR-T/A in HEK293 cells treated with trypsin (F), CS (G), or NE (H) and either vehicle or PAR₂ antagonist (I-343). I, effects of I-343 on PKD activity in the Golgi apparatus stimulated by phorbol 12,13-dibutyrate (PDBu) or thrombin. n = 3–5 experimental replicates, triplicate observations. *, p < 0.05; **, p < 0.01 to vehicle (one-way ANOVA, Bonferroni multiple comparisons). Error bars, S.E.