S. aureus increased CLS1 phosphorylation.To evaluate effects of S. aureus on murine epithelia, lung type II cells were isolated and cultured with the pathogen.
Wild type murine lung fibroblasts (WT ACTN4) and stable ACTN4 knockdown (ACTN4 KD) murine lung fibroblasts were provided by Dr. Pollak's laboratory.
In our studies, the in vivo inhibition of apoptosis in two different cell types, murine lung epithelia and human peripheral blood monocytes, significantly reduced the appearance CD45+Pro-Col-Iα1+ cells, suggesting that this phenotypic change is a non-specific response to local cell death.
As shown in Figure 5, in wild type primary murine lung endothelial cells bearing TSP1, the addition of DEA/NO increased intracellular cGMP production, while treatment with Aβ inhibited DEA/NO stimulated cGMP production, as expected.
The examined category II (mqoB) [61] and III mutants (edd) [60] were attenuated in virulence and caused less inflammation in murine lungs than wild type TBCF10839 (Figure 5).
Re-expression of wild type human ACTN4 in ACTN4 knockdown murine lung fibroblasts reverted cell spreading, cellular and nuclear cross-sectional area, and contractility back towards baseline, demonstrating that the defect was due to absence of ACTN4.
Murine lung epithelial cell line, LA-4, was purchased from American Type Culture Collection (ATCC) (Manassas, VA) and grown in F12K medium containing 15% heat-inactivated fetal bovine serum at 37°C in 5% CO2. Isolation of primary epithelial cells from the lungs of BALB/c and C57Bl/6 mice was performed as previously described [19], with slight modifications.
Longevity assurance homolog 5 (LASS5) was found to be the predominant ceramide synthase in murine primary type II epithelial cells and SV40-transformed murine lung epithelial (MLE) cells.
In order to detect the cellular functions of ACTN4, we tested both wild type ACTN4 (WT ACTN4) and knockdown (ACTN4 KD) murine lung fibroblasts.
Previous analysis revealed that Rab27 proteins are expressed in the murine lung, suggesting that they play an important role in lung cell types with specialized secretory functions (6, 29).
