Journal: Cell Death & Disease
Article Title: Ezrin drives adaptation of monocytes to the inflamed lung microenvironment
doi: 10.1038/s41419-024-07255-8
Figure Lengend Snippet: A Schematic representation of ezrin signaling. Ezrin links plasma membrane (PM) and filamentous actin (F-actin) and adheres to the extracellular matrix (e.g., collagen) via integrin α/β. The activation of focal adhesion kinase leads to efficient cell spreading and adhesion, and PI3K/AKT signaling activation is required for cell survival. B Quantification of crystal violet absorbance emitted from adherent primary BM monocytes of murine WT and Ezr -KO m untreated or treated with LPS (6 h). Absorbance values are relative to WT Untr. C Representative WB of phospho-FAK (pFAK, tyrosine 397), total FAK, phospho-AKT (pAKT, serine 473) and total AKT and densitometric analysis of pFAK and pAKT in primary BM monocytes of murine WT and Ezr -KO m untreated or treated with LPS (6 h). Bar graphs represent pFAK/FAK and pAKT/AKT ratios. Protein fold increase was normalized to β-actin and relative to untreated cells. D Quantification of the percentage of dead cells in primary BM monocytes of murine WT and Ezr -KO m , post-exposure to LPS. See also Fig. . E Relative fluorescence units measuring caspase3/7 activity in primary BM monocytes of murine WT and Ezr -KO m , untreated or treated with LPS at different time points. Data are represented as mean ± SEM from three independent experiments. Statistical analysis was performed using one-way ANOVA or Tukey’s test for multiple comparisons between the genotype and treatment conditions. * p < 0.05, ** p < 0.01 and **** p < 0.001. See also Supplementary Fig. .
Article Snippet: In a set of experiments, BMD-Mφs were plated at the density of 200,000 cells/well (96 well plates) and treated with FAK inhibitor (PND 1186—1 μM, Tocris) or AKT inhibitor (LY 294002—10 μM, Tocris) for 24 h with continued incubation at 37 °C with 5% CO 2 .
Techniques: Membrane, Activation Assay, Fluorescence, Activity Assay
