Figure 3: Ca

<b>(A)</b> Increases in intracellular calcium levels ([Ca²⁺]i) due to external Ca²⁺ entry were demonstrated by augmentation of Fluo-4 signals in stretched AE cells <b>(right panels)</b> but not in control cells <b>(left panels)</b>. Fluorescence recordings were initiated after acute cyclic stretch holding at maximum distension while confocal imaging. <b>(B)</b> Quantitative time curves of Ca²⁺ entry into AE cells in control cells (n = 37), and in cells stretched at 5% (n = 58) and 10% (n = 72) intensity. Bar graph shows mean increase in [Ca²⁺]i at different stretch intensities. <b>(C)</b> Representative time curves of Ca²⁺ entry after 10% stretch in control (n = 45) and stretched AE cells pre-treated with inhibitors of mechanosensitive (GsMTx-4; n = 55), store-operated (2-aminoethoxydiphenyl borate [2-APB]; n = 60) and voltage-gated (nifedipine; n = 55) Ca²⁺ channels. Bar graphs for mean data are shown. <b>(D)</b> Addition of TRPC-6-specific pore-blocking (TRPC-6 Ab) attenuated Ca²⁺ influx in 10% stretched cells (control: n = 40; 10% stretch: n = 36; 10% stretch + TRPC-6 Ab: n = 38). For <b>(B to D)</b>, internal stores were pre-emptied with thapsigargin to rule out contributions from internal endoplasmic reticulum Ca²⁺ release to F/F0. Data are shown as mean ± SEM. *p ❖p &lt; 0.001 versus 10% stretch. See Supplemental Video 1. Abbreviations as in Figures 1 and 2.