Figure 2.

Overexpression of FXYD5 promotes the secretion of inflammatory cytokines by activating the NF-κB signaling in alveolar epithelial cells. (A) MLE-12 cells were incubated with Ad-Null or Ad-FXYD5 as described in Section “Materials and Methods.” The expression of FXYD5 was determined in the CL by Western blot with an HA antibody. (B) A549 cells were treated as in (A) and CL was isolated. Left panel: representative immunoblots. Right panel: densitometric quantification of pIκBα in relation to total IκBα. The expression of FXYD5 was determined with an HA antibody (n = 4). (C) MLE-12 cells were treated as in (A) and CCL2 mRNA was quantified by RT-qPCR (n = 3). (D–E) MLE-12 cells were treated as in (A), culture media was collected, and CCL2, n = 5 (D) and IL-6, n = 5 (E) were determined by ELISA. Values of AdNull-treated controls were normalized to 1. Bars represent means ± SD. Statistical significance was analyzed by unpaired Student’s t-test. *p ≤ 0.05; **p ≤ 0.01.