Fig. 2: Vimentin is phosphorylated by PLK1 at T327, S339, S459, and S83 in vitro. | Cell Death & Differentiation

Fig. 2: Vimentin is phosphorylated by PLK1 at T327, S339, S459, and S83 in vitro.

From: PLK1/vimentin signaling facilitates immune escape by recruiting Smad2/3 to PD-L1 promoter in metastatic lung adenocarcinoma

Fig. 2

a A549 cells were treated with 2.5 ng/ml of TGF-β for 48 hours to induce EMT. Immunoprecipitation of cell lysates was performed with anti-PLK1 antibody or normal IgG followed by immunoblotting with anti-vimentin antibody. b Possible sites on vimentin phosphorylated by PLK1 were newly detected in the LC-MS/MS analysis at the T327, T336, and S339 residues. c Purified GST-tagged wild-type, T327A, T336A, S339A, S459A, and S83A vimentin mutants were used for a PLK1 kinase assay with radioactive ATP. d,e Phosphorylation of vimentin occurs in A549 (d) and NCI-H460 (e) cells treated with 2.5 ng/ml of TGF-β for 48 hours. Treatment with phosphatase (CIP) reduced the phosphorylational modification of vimentin and PLK1 in TGF-β-induced EMT. TCTP was used as a positive control of the PLK1 substrate. All experiments were performed at least three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001; (n = 3). n.s. not significant.

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