Figure 5.

Media utilized for culture maintenance affects levels of spontaneous CFTR activity in non-CF nasal epithelia. Prior to analysis, epithelia were cultured for 96-h in either a proprietary media or a semi-defined media (see “Methods” section for additional details). Epithelia were analyzed in an Ussing chamber using the same methods listed in Fig. 1. (a) Responses to forskolin/IBMX (F/I) and CFTR(inh)-172 in the presence or absence of F/I were quantified. Significant differences due to media were detected for F/I responses [p = 0.0015 for short-circuit current (I_sc)], post-F/I CFTR(inh)-172 responses [p = 0.0205 for transepithelial potential difference/voltage (V_te), p = 0.0017 for I_sc], and spontaneous CFTR activity (p = 0.0277 for V_te). (b) For acutely activated epithelia, the ratio of the change in response to F/I to the change in response to the following CFTR(inh)-172 treatment were calculated. Values shown are absolute. (c) Responses to 172 were normalized to the values obtained in epithelia treated with F/I prior to inhibition. n = 1 donor with 3 replicates per condition.