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. Author manuscript; available in PMC: 2022 Apr 1.
Published in final edited form as: Free Radic Biol Med. 2021 Feb 16;166:90–103. doi: 10.1016/j.freeradbiomed.2021.01.054

Figure 8. Adenoviral overexpression of SIRT6 attenuates menadione-induced Prx hyperoxidation and nuclear accumulation of phosphorylated p65 in human articular chondrocytes.

Figure 8.

Human articular chondrocytes were transduced with an adenoviral vector encoding SIRT6 or an empty vector control for 48 hours and then treated with menadione (25 μM) for 0-30 mins. (A) Cytoplasmic and nuclear fractions were isolated and Prx hyperoxidation was detected by reducing immunoblots with an antibody to PrxSO2/3. Representative immunoblots are shown and specific Prxs are labelled. (B) Densitometric analysis from cytoplasmic and (C) nuclear PrxSO2/3 immunoblots (n=4). Data are expressed as relative intensity compared to untreated empty vector controls. (D) Cytoplasmic and nuclear lysates were also immunoblotted with antibodies to phosphorylated and total p65. LDH (non-nuclear protein) and TBP (nuclear protein) were immunoblotted for, in order to confirm successful cytoplasmic and nuclear separation. Immunoblots are representative results from n=6 independent samples. (E) Densitometric analysis from cytoplasmic and (F) nuclear phospho-p65 immunoblots. Asterisks represent significant differences compared to controls (**, p<0.01; ****, p<0.0001) (Two-way ANOVA). Two-way ANOVA was used to test for differences between time points.