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Fig. 3. Application of high (nN) levels of stress to cell-surface integrins increases intracellular calcium. (A) Phase-contrast view of an adherent endothelial cell with attached RGD-bead (white arrow) bound to integrins on the apical cell surface. Black arrowhead indicates position of the tip of the electromagnet. (B) A time series of pseudocolored fluorescence images of the cell shown in A after mechanical stress (5 nN) was applied with the magnet. These pseudocolored images demonstrate a transient stress-induced increase in [Ca 2+]i as a brief shift in color from blue to yellow, as detected using Fura-2AM ratio-imaging (color bar indicates [Ca 2+]i in nM). The times after start of the time-lapse series are indicated in seconds; the force pulse was applied at 9 seconds in this series. (C) Plot of average [Ca 2+]i for control (
) and gadolinium chloride-treated (
) cells as a function of time; the inhibition of stress-induced calcium influx by gadolinium was statistically significant (P<0.002; error bars indicate s.e.m.). Black arrow indicates when the 3-second force pulse was applied.
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