Fig. 2: Membrane lipid raft disruption causes ligand-free increased dimerization of EGFR. | Nature Communications

Fig. 2: Membrane lipid raft disruption causes ligand-free increased dimerization of EGFR.

From: A Pseudomonas aeruginosa quorum-sensing inducer controls lung permeability in establishing chronic infection via EGFR

Fig. 2: Membrane lipid raft disruption causes ligand-free increased dimerization of EGFR.The alternative text for this image may have been generated using AI.

A 3oc12-induced EGFR phosphorylation in dose depended manner. B Co-cultured supernatants of different strains of P. aeruginosa with A549 cells: ΔlasI strain that lacks the 3oc12 production was less able to induce EGFR phosphorylation. C Fluorescently labeled EGFR-Halo (by JFX646-HaloTag ligand) partially colocalizes with Alexa555-conjugated Cholera toxin-B labeled lipid rafts before, but not after, treatment with 10 mM MβCD or 10 μM 3oc12. Scale bar, 5 μm. The bottom panel showed the fraction of EGFR-Halo spots in lipid raft structures before and after treatment with MβCD or 3oc12. Data are are presented as the mean ± SD (n = 11, 9 and 12 cells). D Stacked Histogram shows fractions of EGFR tracks classified as free, confined, or immobile under 3oc12 stimulation. Data are representative of three independent experiments and are presented as the mean ± SD(n = 32,29). E Distributions of the diffusion coefficient of membrane-docked EGFR molecules before and after 3oc12 treatment. F The diffusion coefficient values for free, confined, and immobile diffusion states of EGFR molecules under 3oc12 stimulation. Data are representative of three independent experiments and are presented as the mean ± SD(n = 32,29). G Confinement radius distribution of confined track segments for each condition. Minimal of 1000 trajectories from approximately 30 cells each in DMSO and 3oc12-stimulated groups from three experiments were selected for analysis(n = 32,28). Data represent mean ± SD. H SMPSC analysis of dimer formation of EGFR-mEGFP on 3oc12-treated cell surface. The results of each group were counted from 10 to 30 cells with more than 4000 individual fluorescent molecules. Data are representative of two independent experiments and are presented as the mean ± SD(n = 12,24,12,24). I Super-resolution structured illumination (SIM) imaging of cortical actin filaments using TIRF illumination mode. The representative images of the actin cytoskeleton of cells with DMSO, 3oc12 (10 μM), or latrunculin B (375 nM) treatment were shown. (C, D, F, G and H). Statistical significance was tested by two-tailed unpaired Student’s t-test. *P < 0.05. **P < 0.01, ****P < 0.0001. Source data are provided as a Source Data file.

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