Figure 2

Electroporation mediated gene delivery of the β1, but not the β2 or β3 subunits, of the Na⁺,K⁺-ATPase attenuates inflammation in LPS injured lungs. Lung injury was established in C57B6 mice (n = 6–8) by aspiration of LPS (5 mg/kg) and 1 day later plasmids (100 μg each) expressing either no insert (pcDNA3), or the β1, β2, or β3 subunits of the Na⁺,K⁺-ATPase (β1, β2, or β3) were delivered in 50 μl to the lungs by aspiration followed immediately by electroporation (8 pulses of 10 ms duration each and 200 V/cm). Naïve mice (n = 5) received no LPS or DNA. Two days after gene transfer (3 days after LPS administration), lungs were lavaged with PBS and BAL fluid was collected and analyzed for cellularity by cytospin followed by Diff-quik staining (A). All experiments were carried out three times and a representative experiment is shown. Total cells were quantified in the BAL fluid and shown as mean ± SEM (B). One-way ANOVA with post-hoc Tukey’s multiple comparisons was used for statistical analysis; a, p < 0.0001 compared to naïve; b, p < 0.05 compared to LPS alone; c, p < 0.05 compared to LPS + pcDNA3; d, p < 0.01 compared to LPS + β1; e, p < 0.05 compared to LPS + β1. The number of PMNs in the BAL fluid were also quantified and shown as mean ± SEM (C). One-way ANOVA with post-hoc Tukey’s multiple comparisons was used for statistical analysis; a, p < 0.0001 compared to naïve; b, p < 0.01 compared to LPS alone; c, p < 0.05 compared to LPS + pcDNA3; d, p < 0.01 compared to LPS + β1; e, p < 0.05 compared to LPS + β1.