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Cyclic stretch induced IL-33 production through HMGB1/TLR-4 signaling pathway in murine respiratory epithelial cells

Fig 5

Cyclic stretch activates TLR4-dependent signaling (NF-κB translocation) and pro-inflammatory state (IL-6 release into medium).

(A) MLE-12 cells were transfected with 50nM TLR-4 specific siRNA or negative control siRNA using Lipofectamine 2000 for 48 h. TLR-4 expression in control group (con), negative control siRNA group (con siRNA) and TLR-4 specific siRNA group (TLR4 siRNA) were detected by western blot and normalized by β-actin (upper graph). (B) Stretch induced NF-κB nucleus translocation were TLR-4 dependent. Total and nuclear NF-κB were measured by Western blot and its nucleus/total NF-κB ratio was analyzed after 6h cyclic stretch in transfected cells. (C) IL-6 secretion in transfected cell media after 6h cyclic stretch was measured by ELISA and was TLR-4 dependent. Each group collected from three wells in a single experiment and the data were presented as mean ± SD from three separate experiments. **P<0.01, ***P<0.001 when compared between groups denoted by horizontal lines.

Fig 5