Fig 1: Treatment with insulin restored HG-induced changes in the HUVEC expression levels of IRSp53 and gal-3. (a) The expression levels of IRSp53 and gal-3 in HUVECs treated with D-glucose at different concentrations and/or 1.5 µg/mL insulin were investigated by Western blotting using ß-actin as a loading control. NG: normal glucose (25 mM); HG: high glucose (60 mM); HG+INS: high glucose (60 mM) + insulin (1.5 µg/mL). The values are the mean ± SD of three independent experiments. ***P < 0.001 vs. the NG group; ###P < 0.001 vs. the HG group.
Fig 2: A high D-glucose concentration affected the expression levels of IRSp53 and gal-3 in HUVECs. (a) The expression levels of IRSp53 and gal-3 in HUVECs treated with D-glucose at different concentration for 96 h were determined by Western blotting using ß-actin as a loading control. NG: normal glucose (25 mM); H1: high glucose 1 (40 mM); H2: high glucose 2 (60 mM); H3: high glucose 3 (80 mM). (b) The expression levels of IRSp53 and gal-3 in HUVECs treated with 60 mM D-glucose for different times were determined by Western blotting using ß-actin as a loading control. (c) The expression levels of IRSp53 and gal-3 in HUVECs treated with 60 mM D-glucose or 60 mM mannitol for 96 h were determined by Western blotting using ß-actin as a loading control. NG: normal glucose (25 mM); HG: high glucose (60 mM); MN: mannitol (60 mM). The values are the mean ± SD of three independent experiments. *P < 0.05 vs. the NG group; **P < 0.01 vs. the NG group; ***P < 0.001 vs. the NG group.
Fig 3: The role of IRSp53 in the expression level of gal-3 and inflammatory state of HUVECs. (a) The expression levels of IRSp53 and gal-3 in normal control (NC) HUVECs, HUVECs transfected with IRSp53 overexpressing lentivirus, and HUVECs transfected with IRSp53-siRNA were detected by Western blotting using ß-actin as the loading control. (b) The expression levels of NF-?B and I?Ba in NC HUVECs and IRSp53-overexpressing HUVECs with different treatments were determined by Western blotting using ß-actin as the loading control. (c) The expression levels of NF-?B and I?Ba in NC HUVECs and IRSp53-knockdown HUVECs with different treatments were investigated by Western blotting using ß-actin as the loading control. (d) Representative images of cellular immunofluorescence staining of NF-?B in HUVECs with different treatments (200x magnification). NC: normal control; OE-IRSp53: overexpressing IRSp53; SI-IRSp53: knockdown IRSp53; normal glucose (25 mM); HG: high glucose (60 mM); HG+INS: high glucose (60 mM) + insulin (1.5 µg/mL). The values are the mean ± SD of three independent experiments. *P < 0.05 vs. the NG group; ***P < 0.001 vs. the NG group; ###P < 0.001 vs. the HG group; ††P < 0.01; †††P < 0.001.
Fig 4: The effect of IRSp53 on HUVEC mobility. (a) Representative images of a scratch assay of HUVECs with different treatments (100x magnification) and the wound closure rate of HUVECs with different treatments. (b) Representative images of a transwell assay of HUVECs with different treatments (200x magnification) and the number of migrated cells. NC: normal control; OE-IRSp53: overexpressing IRSp53; SI-IRSp53: knockdown IRSp53; normal glucose (25 mM); HG: high glucose (60 mM); HG+INS: high glucose (60 mM) + insulin (1.5 µg/mL). The values are the mean ± SD of three independent experiments. ***P < 0.001 vs. the NG group; #P < 0.05 vs. the HG group; ##P < 0.01 vs. the HG group; ###P < 0.001 vs. the HG group; †††P < 0.001.
Fig 5: DM-induced changes in the expression levels of IRSp53 and gal-3 in the aorta of rats. (a) The expression levels of IRSp53 and gal-3 in the aorta of NC rats or DM rats were investigated by Western blotting using ß-actin as a loading control (NC group, n = 6; DM group, n = 10). (b) Representative images of aorta immunofluorescence staining showing levels of IRSp53 and CD31 (20x magnification) (NC group, n = 6; DM group, n = 10); the colocalization ratio of IRSp53 is shown in the bar graph. (c) Representative images of aorta immunofluorescence staining showing levels of gal-3 and CD31 (20x magnification) (NC group, n = 6; DM group, n = 10); the colocalization ratio of IRSp53 is shown in the bar graph. ***P < 0.001 vs. the NC group.
Supplier Page from Abcam for Anti-IRSp53 antibody