Figure 3
From: Sirt1-deficiency causes defective protein quality control
Hsp70-dependent protein quality control is impaired in Sirt1–/– MEFs.
(A) Immunoblotting of lysates from wild-type and Sirt1–/– MEFs. (B) Degradation of pVHL was monitored by immunoblotting. HA-tagged pVHL was transfected into wild-type and Sirt1–/– MEFs and 100 μg/ml cycloheximide (CHX) was added prior to harvest at the indicated times. Actin serves as a loading control. (C) Cycloheximide chase as described in (B) was performed in wild-type, Sirt1–/–, Sirt1–/– transfected with wild-type Sirt1 or Hsp70 and wild-type MEFs treated with 10 μM MG132. Protein levels of HA-pVHL were quantified. Actin was used for normalization. The data represent means ± SEM from three independent experiments. Uncropped gel images are shown in Supplementary Figure.
