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. Author manuscript; available in PMC: 2021 Jun 10.
Published in final edited form as: Curr Res Toxicol. 2020 Sep 24;1:111–115. doi: 10.1016/j.crtox.2020.09.003

Figure 1: Effects of KIF and AUY-922 on lung endothelial cells.

Figure 1:

Western Blot analysis of (A) phosphorylated Cofilin (pCofilin) and Cofilin (B) phosphorylated MLC2 (pMLC2) and MLC2 after 24 hours treatment of BPAEC with either KIF (5 μM) or vehicle (0.1% DMSO) and post-treatment with AUY-922 (2 μM) or vehicle (0.1% DMSO) for 16 hours. The blots shown are representative of 4 independent experiments. The signal intensity of the bands was analyzed by densitometry. Protein levels of pCofilin and pMLC2 were normalized to Cofilin and MLC2 respectively. *P < 0.05, **P<0.01 vs vehicle. Means ± SEM. Western Blot analysis of (C) phosphorylated MLC2 (pMLC2) and MLC2 after 24 hours treatment of HuLEC with either KIF (5 μM) or vehicle (0.1% DMSO); and post-treatment with AUY-922 (2 μM) or vehicle (0.1% DMSO) for 16 hours. The blots shown are representative of 3 independent experiments. The signal intensity of the pMLC2 was analyzed by densitometry. Protein levels of pMLC2 were normalized to MLC2. *P<0.05 vs vehicle. Means ± SEM. (D) Confluent monolayers of BPAEC were pre-treated with either vehicle (0.1% DMSO) or KIF (25 μM) (red arrow) for 18 hours, followed by treatment with either vehicle (0.1% DMSO) or 5 μM of AUY-922 (green arrow). A gradual increase in endothelial permeability (reduced TEER) was observed in KIF treated cells (red line). AUY-922 significantly reduced the endothelial permeability (increased TEER) in both KIF-pretreated (blue line) and vehicle-treated cells (green line). (E) Cells were incubated with either VEH (0.1% DMSO) or KIF (0.01, 0.1, 1, 10, 100, 200 μM) or AUY-922 (0.01, 0.1, 1, 10, 100, 200 μM) for 24 hours. Cellular viability was evaluated by employing the MTT assay. ***P<0.001 vs VEH, n=3. Means ± SEM.