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. 2018 Mar 27;114(6):1455–1466. doi: 10.1016/j.bpj.2018.01.027

Figure 5.

Figure 5

Frequencies of calcium spike and MT1-MMP vesicle delivery are biphasically regulated by ECM cross-linking. (A) Shown are representative fluorescence micrographs of Cerulean-cortactin-MTLn3 (cyan, middle) cells (see Movie S6) labeled with Fluo-4-AM (green, top), plated on fluorescent gelatin (red, bottom). Scale bars,10 microns. (B) Representative calcium spikes are shown, recorded in cells at different gelatin cross-linking ratios. (C) Shown is a power spectrum of calcium spikes in (B). (D) Shown is a comparison of average spike frequencies at different gelatin cross-linking ratios. Scale bars, 10 μm. Data include >15 cells per experiment for three separate experiments per condition. Pairwise comparisons were done with a 0.39 cross-linking ratio condition. (E) Shown is a representative image of an HS-578T cell transfected with MT1-MMPpHluorin (see Movie S6). The right panel depicts the dynamics of MT1-MMP containing vesicle delivery to the plasma membrane. The red arrows point at newly delivered vesicles. (F) Shown is the representative record of the MT1-MMP delivery event over 30 min in cells at different gelatin cross-linking ratios. (G) A comparison of the average frequency of MT1-MMP delivery is shown at different gelatin cross-linking ratios. Data include >15 cells per experiment for three separate experiments per condition. Pairwise comparisons were done with a 0.39 cross-linking ratio condition. Errors are shown as mean ± SE. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. To see this figure in color, go online.