Fig 1: Sesn2 levels in aortic valves from patients with CAVD. A and B, Sesn2 expression in normal donors and patients with CAVD was examined by immunofluorescence staining (×400) and western blotting. C, Double immunofluorescence staining with anti-Sesn2 and anti-F4/80 antibodies (×400). N = 10 for the normal group and 20 for the CAVD group. *P < 0.05 versus the normal group.
Fig 2: Effect of Sesn2 on M1 macrophage-related oxidative stress in vitro. A, Nrf2 levels in each group were measured by western blotting. B, ROS levels were examined using DCFH-DA. C, SOD activity, MDA levels, and GSH levels were analyzed. N = 6 in each group. *P < 0.05 versus the ox-LDL + siRNA or cDNA-Sesn2 group. #P < 0.05 versus the ox-LDL + si-Sesn2 or cDNA-Sesn2 group.
Fig 3: Effects of the Nrf2 pathway on osteoblastic differentiation of VICs. The mRNA expression levels of ALP, Runx2, and OPN in VICs were measured. N = 6 in each group. *P < 0.05 versus the VICs group. #P < 0.05 versus the ox-LDL + VICs group. &P < 0.05 versus the ox-LDL + Mø group. ?P < 0.05 versus the ox-LDL + Mø + si-Sesn2 + ML385 group.
Fig 4: iNOS mRNA levels and expression of osteoblastic differentiation markers (VICs) in CAVD. A, The mRNA expression levels in the 2 groups were examined by RT–qPCR. B, Correlation between iNOS mRNA levels and Sesn2, ALP, Runx2, and OPN mRNA levels. N = 10 for the normal group and 20 for the CAVD group. *P < 0.05 versus the normal group.
Fig 5: Effect of Sesn2 on ox-LDL–induced M1 macrophage polarization in vitro. A, F4/80+CD86+ cells were examined in the 5 groups by flow cytometry. B, CD86 intensity was measured using immunofluorescence staining. C, The mRNA expression levels of CD38, CD80, CD86, and iNOS were investigated by RT–PCR. N = 6 in each group. *P < 0.05 versus the ox-LDL + siRNA or cDNA group.
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