Skip to main content
. 2021 Aug 3;48(4):185. doi: 10.3892/ijmm.2021.5018

Figure 4.

Figure 4

In vitro blocking of VCAM-1 suppresses neutrophil adhesion and migration through Transwells. mGECs from the HO-1+/− and wild-type mice were isolated. (A) Western blot analysis of the expression levels of HO-1, VCAM-1 and β-actin proteins in the mGECs extracted from the HO-1+/+ and HO-1+/− mice. (B) Immunofluorescence staining of VCAM-1 in the mGECs extracted from the HO-1+/+ and HO-1+/− mice. Relative fluorescent intensity was quantified using ImageJ software (n=3, *P<0.05 vs. HO-1+/+). (C) mGECs were grown in a 96-well plate or Transwell chamber and stimulated with 100 U/ml TNF-α for 4 h. Neutrophils were isolated and labeled with PKH26 to perform adhesion assay or Transwell migration assay. (D) Neutrophils adhered to mGECs were photographed using a fluorescence microscope, and the fluorescence area was quantified using ImageJ software (n=3, *P<0.05 vs. HO-1+/+; #P<0.05 vs. HO-1+/−). (E) Neutrophil migration through mGECs was photographed using a fluorescence microscope, and the fluorescence area was quantified using ImageJ software (n=3, *P<0.05 vs. HO-1+/+; #P<0.05 vs. HO-1+/−). mGECs, mouse glomerular endothelial cells; HO-1, heme oxygenase-1; VCAM-1, vascular cell adhesion molecule-1; Ab, antibody.