Mediators of Inflammation / 2021 / Article / Fig 6

Research Article

Ataxin-10 Inhibits TNF-α-Induced Endothelial Inflammation via Suppressing Interferon Regulatory Factor-1

Figure 6

Ataxin-10 binds to and inhibits IRF-1 to suppress TNF-α-induced VCAM-1 in HUVECs. (a) Cell lysates were harvested from Y2HGold yeast strain transfected with pGBKT7 or pGBKT7-Ataxin-10. The expression of Ataxin-10 was detected by Western blotting. (b) HUVECs were treated with 10 ng/mL TNF-α for 2 hours. Immunoprecipitation experiment was performed with Ataxin-10 antibody and IgG (control), showing specific interaction between Ataxin-10 and IRF-1 in HUVECs. (c) Western blotting analysis of endogenous IRF-1 subcellular localization in HUVECs transfected with empty vector of HA-Ataxin-10. The purity and equal loading of cytoplasmic fractions and nuclear fractions were confirmed by tubulin and TBP, respectively. (d) HUVECs were transfected with Ataxin-10 siRNA or IRF-1 siRNA or both and then treated with TNF-α (10 ng/mL) for 8 hours. VCAM-1 and ICAM-1 protein levels were detected by Western blotting.

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