Fig. 1.
Sirtuin 7 activity is not only involved in the regulation of rDNA transcription. (A) In vitro sirtuin 7 deacetylase assays using fluorescent H3K18-Ac peptides performed with or without preincubation of full-length recombinant human sirtuin 7 with sirtinol or EX-527. Results represent mean±s.d. (n=3). ***P<0.001 compared with control (ANOVA and Dunnett's t-test). (B) Asynchronous HeLa cells were pulse labeled with [32P]orthophosphate for 2 h and chased in nonradioactive medium with or without sirtinol for various times up to 6 h. Total RNAs were isolated, resolved on the same 1% agarose formaldehyde gel and detected by ethidium bromide staining. The RNAs were then transferred to a positively charged membrane and UV cross-linked. (a) Autoradiography of newly synthesized RNAs before chase (lane 1) and after increasing times of chase in the absence (lanes 2–6) and in the presence of sirtinol (lanes 7–11). (b) The upper part of the autoradiography (asterisk) after applying a global intensity adjustment. (c) Detection of RNAs loaded on the gel by ethidium bromide staining.