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. 2023 Nov 20;14:1277582. doi: 10.3389/fimmu.2023.1277582

Figure 2.

Figure 2

rCC16 treatment increases SPLUNC1 expression in vitro and in vivo. (A) WT (n=3) and CC16-/- (n=3) MTECs were treated with media (control) or rCC16 (25μg/ml) for 24 hours, after which BPIFA1 expression in cell lysates was measured by RT-PCR with GAPDH as a housekeeping control. *P<0.05, * *P<0.01, * **P<0.001, * ***P<0.0001 by One-Way ANOVA Tukey’s multiple comparison test. SPLUNC1 apical protein expression was confirmed for control- and rCC16-treated WT (B) and CC16-/- (C) MTECs by western blotting (n=3 per group). The same volume of protein (20μl) was loaded for each sample. **P<0.01, ***P<0.001 by Unpaired t test. Data are presented as mean±SEM. (D) CC16-/- mice were treated intravenously with saline (n=10) and rCC16 (n=10) for 3 days, after which BPIFA1 expression in lung tissue was measured by RT-PCR with GAPDH as a housekeeping control. *P<0.05 by Unpaired t test. (E) SPLUNC1 protein expression was measured in the BALF from the 3-day saline (n=4) and rCC16 (n=4) treated CC16-/- mice. The same volume of protein (20μl) was loaded for each sample. **P<0.01 by Unpaired t test. Data are presented as mean±SEM.