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Fig. 8. Myofibroblasts and fibroblasts exhibit different kinetics of intercellular Ca²⁺_[i] wave propagation. Myofibroblasts (A-C) and fibroblasts (D-F) were grown between 50-µm-wide non-adhesive lines, created on glass by means of microcontact printing (A,D, Fura-2 Em₃₈₀ fluorescence image, scale bar: 25 µm) (see also supplementary material Movie 3). Individual cells were locally stimulated by touching gently with a micropipette (*); Ca²⁺_[i] transients were analysed in the stimulated cell (yellow outline) and all following cells in the chain (blue to red to green; grey traces depict non-contacting control cells). Fura-2 Em₃₄₀/Em₃₈₀ fluorescence ratios were recorded at 1 frame/second over the indicated cell outlines and plotted over time with the respective colour (B,E); arrows indicate time delay of Ca²⁺_[i] transient initiation after stimulation of the first cell (black line). A slow fluid flow applied perpendicularly to the lines excluded the possibility that released soluble factors induce a Ca²⁺ response. For quantitative analysis, fluorescence ratio intensity values were recorded along a line drawn across all cells in the chain (dotted line; A, 250 µm; D, 175 µm) and are displayed over time in a kymograph. In the kymograph image, delays of Ca²⁺_[i] wave propagation at cell junctions appear as clear steps along the time axis (t=40 seconds) in the case of myofibroblasts (B) but are hardly detectable between fibroblasts (E). All time delays at cell junctions were measured on kymograph images and summarised in histograms for myofibroblasts (C, n_exp=17, n_pairs=34) and for fibroblasts (F, n_exp=24, n_pairs=55).

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