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. 2022 May 18;9:792717. doi: 10.3389/fcvm.2022.792717

FIGURE 4.

FIGURE 4

OxLDL-induced vimentin (Ser72) phosphorylation depended on CD36 and led to disassembly of vimentin filaments. (A) Western blots for phosphorylated vimentin (Ser72) and phosphorylated vimentin (Ser38) using cell lysates of wild type murine peritoneal macrophages. Cells were treated with oxLDL (50 μg/ml) for indicated times. (B) Western blot for phosphorylated vimentin (Ser72) using cell lysates of wild type murine peritoneal macrophages. Cells were treated with various concentrations of oxLDL for 10 min. (C) Wild type murine peritoneal macrophages were incubated with OxLDL (50 μg/ml) for indicated times. Cytosolic and membrane fractions were separated using buffer-based protocol. E-cadherin was used as a marker for plasma membrane fraction and beta actin was used as a marker for cytosolic fraction. (D) Wild type and CD36 null murine peritoneal macrophages were incubated with myeloperoxidase (MPO)-modified LDL (oxLDL, 50 μg/ml) for indicated times. The lysates were analyzed by western blot for phosphorylated vimentin (Ser72). (E) Macrophages were treated with oxLDL (50 μg/ml) for 10 min. Cytosolic fraction is divided based on the solubility in triton X-100. Western blot for vimentin was done. GAPDH was used as a marker for soluble fraction and flotilin-1 was used as a marker for insoluble fraction. ***p < 0.001. The graph shows mean ± SEM for triplicated determinants of the experiments.