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. 2012 Dec;47(6):729–737. doi: 10.1165/rcmb.2012-0057OC

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Copyright © 2012 by the American Thoracic Society

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Figure 3. — Evaluation of the role of phosphodiesterase (PDE)4 in mediating attenuation of acute PGE2 signaling after 24-hour PGE2 pretreatment. (a) Detection of PDE4 isozyme levels after PGE2 pretreatment (24 h) in HFL cells by immunoblot. Black arrows point to a predicted band based on the documented molecular weight of specific PDE4 isozymes. (b) Effect of inhibition of PDE4 with roflumilast (rof, 1.0 μM) and rolipram (rol 1.0 μM) on PGE2-mediated inhibition of HFL chemotaxis after 24-hour PGE2 pretreatment. (c) Effect of inhibition of PDE4 with rof on PGE2-mediated cAMP accumulation after 24-hour PGE2 pretreatment. (d) Effect of inhibition of PDE4 with rol on PGE2-mediated inhibition of HFL chemotaxis after 24-hour PGE2 pretreatment. (e) Time-dependent attenuation of PGE2-mediated cAMP accumulation in HFL cells begins within 15 minutes and is complete by 90 minutes. The addition of roflumilast at 30 minutes restores cAMP accumulation. Cells pretreated with PGE2 do not accumulate cAMP until treated with roflumilast. *P < 0.05; **P < 0.01. Bars are means ± SEM; n = 3 separate experiments in all panels unless otherwise stated.