Figure 6
Effect of TPPP/p25 and/or chemical inhibitors on the SIRT2 activity on tubulin. (a) Inhibition of deacetylation of 2.64 µM tubulin (black column) by 20 µM TPPP/p25 or chemical compounds; 0.6 µM SIRT2 was added to the acetyl-tubulin. The data are presented as mean ± SD, n = 3–11. *p = 5.56E-06 for TPPP/p25, *p = 1.53E-03 for SH1, *p = 1.38E-03 for MZ242 and *p = 1.60E-03 for 1 mM nicotinamide when the values with and without inhibitor were compared in the case of tubulin as a substrate (two-sided, unpaired Student’s t-test). Images of the full-length blots/gels are presented in Supplementary Fig. S5. (b) Effect of the inhibitors on the binding of 125 nM TPPP/p25 to the immobilized SIRT2 detected by ELISA as described in the Materials and Methods. The data are presented as mean ± SD, n = 3. (c) Mutual effect of the TPPP/p25 and the chemical inhibitors on the SIRT2 activity (Western blot data with tubulin). The data are presented as mean ± SD. n = 4 for tubulin, tubulin with SIRT2, tubulin with SIRT2 and 10 μM TPPP/p25, tubulin with SIRT2 and 5 μM MZ242, and tubulin with SIRT2 and 10 μM TPPP/p25 and 5 μM MZ242; n = 3 for tubulin with SIRT2 and 5 μM SH1, and tubulin with SIRT2 and 10 μM TPPP/p25 and 5 μM SH1. *p = 1.06E-05 for TPPP/p25, *p = 4.96E-03 for MZ242 and *p = 1.38E-03 for SH1 when the values with and without inhibitor were compared (two-sided, unpaired Student’s t-test). Images of the full-length blots/gels are presented in Supplementary Fig. S6.
