Fig. 5. Effect of rhHAPLN1 on ROS levels and cell survival in HUVECs. (A) Young HUVECs were treated with rhHAPLN1 and then treated with H2O2. Cells were washed and stained with H2DCFDA. Senescent HUVECs were treated with rhHAPLN1, cells were washed and stained with H2DCFDA. The cellular ROS levels measured by flow cytometry (FACS). (B) The mRNA levels of IL-1β, CCL2, and IL-6 between the groups treated with IL-17A and rhHAPLN1 were determined by qRT-PCR. (C) Senescent HUVECs were treated with rhHAPLN1. The mRNA levels of IL-17A, IL-1β, CCL2, and IL-6 were determined by qRT-PCR. (D, E) The protein levels of eNOS and p-eNOS in the various groups were determined by western blot analysis. #p<0.05; ##p<0.005; and ###p<0.0005 versus Control (young HUVECs). **p<0.005; and ***p<0.0005 versus H2O2 treatment or IL-17A treatment or Control (senescent HUVECs, n=3 per group).
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