Additional file 4: Figure S3. of Heparan sulfate proteoglycans mediate Aβ-induced oxidative stress and hypercontractility in cultured vascular smooth muscle cells

HSPG mitigates Aβ1-40-induced mitochondrial and cytosolic ROS production in VSMC under physiological oxygen concentration. To determine if differing levels oxygen impact ROS production in Aβ1-40 treated VSMC, cells were kept in 10 % oxygen (Panel A) or 1 % oxygen (conditions that are considered hypoxic; Panel B) in cell culture incubator with % 5 CO2. Primary human cerebral VSMC were pre-treated with heparin (15 U/mL), heparinase I (HpnI; 5 Sigma U/mL), or heparinase III (HpnIII; 2 Sigma U/mL) for 2 h, washed, loaded with Mitotracker Red CM-H2XRos, washed, and treated with Aβ1-40. In some cases, cells were pre-treated with heat-inactivated (HI) enzyme. Fluorescence was measured after 30 minutes. Results are representative of 3 independent experiments performed in triplicate. *p < 0.05 vs. vehicle-treated control. #p < 0.05 vs. comparison group. (JPEG 70 kb