Figure 3. C-Src mediated TLR4-dependent EGFR transactivation in MPMs exposed to LPS.

(A–D) MPMs were pre-treated with 10 μM TAK242 or 10 μM PP2 or vehicle (DMSO) for 30 min prior to stimulation with LPS (1 μg·mL⁻¹) for 30 min. The total protein was collected and the levels of p-EGFR, p-c-Src, TLR4, and GAPDH were detected by Western Blot assay. (E) The TLR4 protein level was detected in MPMs acquired from TLR4^−/− mice. (F and G) MPMs were extracted from WT mice and TLR4^−/− mice, respectively. WT MPMs and TLR4^−/− MPMs were treated with LPS (1 μg·mL⁻¹) for 30 min to detect the EGFR phosphorylation level using Western Blot assay. (H and I) MPMs were extracted from WT mice and TLR4^−/−mice, respectively. WT MPMs and TLR4^−/− MPMs were treated with LPS (1 μg·mL⁻¹) for 5 min to detect the level of p-EGFR/p-c-Src complex using immunoprecipitation assay. Each experiment was detected at least three times. Bars represent the mean ± SEM of more than three independent experiments performed in duplicate, and asterisks indicate significant inhibition (*p < 0.05, **p < 0.01, vs. LPS group).