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. 2016 Oct 24;113(45):12715–12720. doi: 10.1073/pnas.1609227113

Fig. S6.

Fig. S6.

Generation of functional stable cell lines Sirt1−/−/SIRT1-EGFP and Sirt1−/−/SIRT1(H355A)-EGFP. (A) Confocal images showing the location of the stable transfected nuclear SIRT1-EGFP and the catalytically inactive SIRT1(H355A)-EGFP together with the transiently transfected nucleolar marker mOrange2-Fibrillarin fusion protein. (B) Western blot from total protein extracts from 1, WT MEFs; 2, Sirt1−/− MEFs; 3, Sirt1−/−/SIRT1-EGFP MEFs; and 4, Sirt1−/−/SIRT1(H355A)-EGFP. Extracts were blotted against the indicated antibodies. (C) RT-PCR analyses from total RNA harvested at the indicated time points after synchronization using dexamethasone (DEX) shock. The amount of Dbp and Per2 mRNA was determined by real time PCR. Means ± SEM of three independent experiments are presented.