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. 2021 Jun 8;16(6):e0252816. doi: 10.1371/journal.pone.0252816

Fig 5. SIRT1/FOXO3a complex directly interacts with the IL-10 promoter region to induce its expression.

Fig 5

(A) Immunoblot of IL-10 expression levels treated with VitD3 in SIRT1 knocking down HAECs (upper) or FOXO3a knocking down HAECs (bottom). Scramble shRNA transfected as control. (B) qPCR to quantify the mRNA levels of IL-10 on the VitD3 treated SIRT1 knocking down HAECs or FOXO3a knocking down HAECs (**p < 0.01, n = 3). (C) Genomic analysis of the promoter region and two traditional FOXO3a binding domains were acquired on -701 to -694 (TGGAAACA) and -208 to -201 (TGAAAACT). Gene modification to delete either of these two fragments and named as IL-10-Del1 or IL-10-Del2. (D) Luciferase reporter assay performed with IL-10-wt, IL-10-Del1 or IL-10-Del2 under VitD3 treatment (**p < 0.01, n = 3). (E) CHIP assay performed with the FOXO3a antibody under VitD3 treatment of HAECs.