Figure 3.

Quorum sensing inhibition with HDMF counteracts the negative effect of P. aeruginosa on CFTR expression, localization and function. PAO1 bacteria were grown in LB with or without 0.125 mg/ml HDMF prior to collection of the bacterial filtrates. CFBE-wt cells were then treated for 24 h with exoproducts from PAO1 cultured with (PAO1+HDMF) and without HDMF (PAO1). LB medium was used as control. (A) Representative immunoblots (left) and densitometric analysis (right, reported as % of LB control) of mature (band C) CFTR protein expression levels (n = 6). (B) Representative immunofluorescence images (of n = 4 independent experiments) of CFBE-wt cells treated with LB, PAO1, or PAO1+HDMF exoproducts. CFTR was detected with an anti-CFTR antibody (Ab) coupled to Alexa Fluor 488 conjugated anti-mouse antibody. Nuclei were stained with DAPI (blue). (C) Representative traces of short-circuit current (I_sc) measurements in Ussing chamber and quantification of mean CFTR_Inh172 (20 μM)-sensitive currents (ΔI_{CFTRInh−172}) through CFBE-wt cells, in each condition (n = 10). NS, non-significant, ^*p < 0.05, ^**p < 0.01, ^***p < 0.001.