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ramp

ramp + hold. control. step. ramp. p-ERK. ERK. step. ramp. ramp + hold. t = 0’. 30’. 60’. PTX. a. b. - +. - +. Pressure. p-ERK. ERK. Apyrase. - +. - +. Pressure. p-ERK. ERK.

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ramp

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  1. ramp + hold control step ramp p-ERK ERK step ramp ramp + hold t = 0’ 30’ 60’ PTX a b - + - + Pressure p-ERK ERK Apyrase - + - + Pressure p-ERK ERK Supplementary Figure 1. A.NHBE cells were exposed to compressive stress using either a 30 minute step increase to a constant pressure, a ramp increase in pressure applied gradually over 30 minutes, or a ramp and hold pattern applied gradually over 30 minutes and held constant for 30 minutes. ERK phosphorylation was evaluated for each loading paradigm by Western blotting with phosphospecific antibodies, and compared to no stress control. B. NHBE cells were pretreated from the basal medium with pertussis toxin (100 ng/ml) or from both apical and basal surfaces with the ATP/UTP scavenger apyrase (20 U/ml). Neither treatment attenuated the compression-induced ERK phosphorylation at 30 minutes, as measured by Western blotting (blots are representative of two experiments each).

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