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. 2014 Jul 25;171(16):3814–3826. doi: 10.1111/bph.12738

Figure 6.

Figure 6

Generation of PAR2-activating proteases. Extracts (30 μg protein) from tissues treated with PAR2-AP (50 μg), bradykinin (5 μg) or NaCl (0.9%) were bioassayed for effects on [Ca2+]i measured in single cells. (A–D) show representative changes in [Ca2+]i in KNRK-mPAR2 and KNRK-VC cells, with each line a response of a single cell. Extracts from tissues pretreated with PAR2-AP (A) or bradykinin (BK) (C) rapidly increased [Ca2+]i in KNRK-mPAR2 cells. This effect was not detected in KNRK-VC cells (B). Extracts from tissues treated with NaCl did not rapidly raise [Ca2+]i in KNRK-mPAR2 cells (D). (E) shows the magnitude of the peak increase in [Ca2+]i in KNRK-mPAR2 or KNRK-VC cells. Melagatran reduced or prevented the increase in [Ca2+]i of KNRK-mPAR2 cells challenged with extracts of tissue from PAR2-AP- and bradykinin-treated mice. n = 2–5 coverslips per group, with 16–36 cells analysed. Extracts were assayed from n = 4 mice per experimental treatment. *P < 0.05 compared with NaCl-treated tissues in KNRK-PAR2 cells. #P < 0.05 vehicle compared with melagatran, bradykinin-treated tissues in KNRK-PAR2 cells. **P < 0.05 responses in KNRK-VC compared with KNRK-PAR2 cells.