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Injury induces an intercellular Ca²⁺ wave. HCE-Ts were loaded with 4 mM fluo-3 AM for 30 minutes and washed two times. Images were captured every 789 milliseconds using a Zeiss LSM 510 Axiovert confocal laser scanning microscope equipped with an Argon laser. A circular wound 250 mm in diameter was made after scanning the cells for 76 seconds. An immediate elevation in intracellular Ca²⁺ begins first along the wound edge and then quickly spreads to neighboring cells in a radial pattern. The response diminished to background levels within two minutes. Movie speed is three frames per second (2.4 X real time).

The injury-induced Ca²⁺ wave is not inhibited in the absence of extracellular Ca²⁺. HCE-Ts were incubated in Ca²⁺-free HEPES-buffered saline containing 4 mM fluo-3 AM for 30 minutes and washed two times. Images were captured using a Zeiss LSM 510 Axiovert confocal laser scanning microscope equipped with an Argon laser. Absence of extracellular Ca²⁺ does not inhibit propagation of the wave but prevents an elevation in [Ca²⁺]_i for cells immediately adjacent to the injury site. Movie speed is three frames per second (2.4 X real time). (See Fig. 6B).

The injury-induced Ca²⁺ wave is inhibited in the presence of thapsigargin. HCE-Ts were incubated in HEPES-buffered saline containing 1 mM thapsigargin and 4 mM fluo-3 AM for 30 minutes and washed two times. Images were captured using a Zeiss LSM 510 Axiovert confocal laser scanning microscope equipped with an Argon laser. Cells treated with 1 mM thapsigargin do not exhibit propagation of a Ca²⁺ wave upon injury and only cells located immediately adjacent to the injury site display an elevation in [Ca²⁺]_i. Movie speed is three frames per second (2.4 X real time). (See Fig. 6C).

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