Fig. 4

Effects of sevoflurane on junction proteins and RhoA/pMLC/F-actin pathway of mouse lung epithelial (MLE-12) cells. Western blots of a ZO-1 and E-cadherin, b total myosin light chain (MLC) and phosphorylated myosin light chain 2 (Ser19) (pMLC2) levels at 6 h in untreated MLE-12 cells (Medium) and cells exposed to sevoflurane alone (Sevo), cytomix alone (Cyto) or cytomix and sevoflurane (Cyto + Sevo). c–e Protein expression levels were quantified and standardized by GAPDH protein level, and pMLC levels were standardized by total MLC levels, expressed as ratios to those in the Medium group. f RhoA activity was standardized by total RhoA protein level at 30 min in identical conditions. All results are reported as medians with interquartile ranges. One-way ANOVA was performed, with post-hoc comparisons if ANOVA results showed significance (compared to the Medium group: *p < 0.05; **p < 0.01; compared to the Cyto group: ^##p < 0.01).g Immunostaining after 6 h of treatment of pMLC and F-actin was performed in identical conditions. Cells were fixed, permeabilized, and stained with antibodies, followed by A488 secondary antibodies and Hoechst. All images were acquired by fluorescent microscope with a 40× objective. Scale bar: 50 μm