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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 4. Effect of adenoviral overexpression of SIRT6 on basal TXNIP, Prx1 and Srx levels in normal human articular chondrocytes.

Figure 4.

(A) Human articular chondrocytes were transduced with an empty adenoviral vector control (ad-Null) or an adenoviral vector encoding SIRT6 (ad-SIRT6) for 48 hours. Basal protein levels of TXNIP, Prx1 and Srx from total cell lysates were assessed by immunoblotting. Representative results from two separate individuals are shown in panels A, D, and F. (B) Densitometric analysis from TXNIP (n=18), Prx1 (n=21) and Srx (n=7) immunoblots. Total protein levels were normalized to β-tubulin as a loading control. (C) Chondrocytes were transduced with ad-Null or ad-SIRT6 for 48 hours and basal TXNIP, Prx1 and Srx mRNA expression were quantified by RT-qPCR (n=6 independent samples). Human articular chondrocytes were transduced with ad-SIRT6 or ad-Null for 48 hours and nuclear fractions (D,E) and cytoplasmic fractions (F,G) were isolated. Basal TXNIP (n=9), Prx1 (n=9), and Srx (n=6) were assessed by immunoblot. Nuclear protein levels were normalized to TBP and cytoplasmic protein levels were normalized to LDH. In all cases, representative immunoblots are shown. SIRT6 immunoblots confirmed SIRT6 overexpression. Paired t-tests were used to test for statistical differences between groups.