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. 2022 Jan 4;132(1):e150051. doi: 10.1172/JCI150051

Figure 4. SIRT6 deacetylates Runx2.

Figure 4

(A) Representative IF images showing the colocalization of SIRT6 and Runx2. Scale bars: 50 μm. (B) Anti-SIRT6 IP followed by Western blot with anti-Runx2 or anti-SIRT6 antibody in SIRT6-Tg VSMCs after treatment with Pi (3.0 mM) for 7days. Anti-rabbit IgG IP was used as a negative control. (C) Anti-Runx2 IP in SIRT6-Tg VSMCs after treatment with Pi (3.0 mM) for 7days. Western blot was carried out with anti-SIRT6 or anti-Runx2 antibody. Anti-mouse IgG IP was used as a negative control. (D) The anti-HA IP and anti- flag IP followed by Western blot with anti-HA or anti-flag antibody in HEK-293T cells infected with HA-Runx2 plasmid, flag-SIRT6 plasmid, or both. Anti-rabbit IgG IP was used as a negative control. (E) WT and SIRT6-Tg VSMC lysates were immunoprecipitated with anti-Runx2 antibody and immunoblotted with anti-acetylated lysine antibody. (F) HEK-293T cells were infected with HA-Runx2 plasmid, flag-SIRT6 plasmid, or both. The anti-HA IP followed by Western blot with anti-acetylated lysine antibody and anti-HA antibody. (G) SIRT6-Tg VSMCs were pretransfected with siSIRT6 or siNC together with Pi (3.0 mM) for 7 days and OSS-128167 or DMSO were incubated with Pi (3.0 mM) for 7 days. The cell lysates were immunoprecipitated with anti-Runx2 antibody and immunoblotted with anti-acetylated lysine antibody and anti-Runx2 antibody. All the above experimental processing were duplicated 3 times.