Co-localisation of thrombin-induced translocation of PKCε with other neuronal markers. A. PKCε translocation (green) following exposure to thrombin (100 nM, 30 s) colocalises with other neuronal markers as shown. PKCε translocation was co-localised in c. half of cells with expression of the neuropeptide CGRP and with the neurofilament marker N52, and in a smaller proportion of cells with the neuropeptide substance P (SP) (panels on right). PKCε translocation was not in general co-localised with IB4 binding nor with parvalbumin (Prv) or COX-1 (panels on left). Neurones from adult mice cultured in the absence of NGF, with the exception of the COX-1 experiment which was carried out in neonatal rat sensory neurons cultured in NGF (100 ng/ml) as the antibody available to us did not bind mouse COX-1. Scale bars all 5 μm.
B. Summary of results from experiments similar to those shown in A. First bar shows percentage of cells showing translocation of PKCε in response to thrombin (100 nM, 30 s). Remaining bars show percentages of these thrombin-responsive cells which co-expressed the neuronal markers noted beneath each bar. White bar in N52 column shows the proportion of the N52 positive neurons in which TRPV1 expression had been demonstrated by recording a calcium increase in response to capsaicin prior to fixation (c.f. Fig. 2). Final white bar shows overall fraction of thrombin-responsive neurons in which TRPV1 expression had been demonstrated by calcium imaging.