Journal: Cells

Article Title: A Rise in ATP, ROS, and Mitochondrial Content upon Glucose Withdrawal Correlates with a Dysregulated Mitochondria Turnover Mediated by the Activation of the Protein Deacetylase SIRT1

doi: 10.3390/cells8010011

Figure Lengend Snippet: The increase in mitochondrial ATP production involves the increased content of mitochondria. ( A ) Cells transfected with siRNA targeting Mfn1 were incubated in medium lacking glucose for 72 h. Mitochondria were immunostained with anti-Tom20 antibody and visualized. The Mfn1 blot (right) shows decreased Mfn1 protein level in cells treated with siMfn1 RNA. All scale bars in microscopy indicate 5 μm; ( B ) cells were glucose deprived for three days, and mitochondrial content in cells treated with siCtrl (black bar) or siMfn1 (grey bar) was measured by staining with NAO and flow cytometric analysis. All flow cytometry was carried out in, at least two biological repeats; ( C , D ) total ATP level, or the amount of ATP produced by oxidative phosphorylation in cells treated with siCtrl (black bar) or siMfn1 (grey bar). Cells were glucose deprived for three days. Values are presented as mean ± s.d.; * p < 0.05, ** p < 0.01 by ANOVA.

Article Snippet: Cells were transfected with small interfering RNA (siRNA) corresponding to human SIRT1 or Mfn1, or control RNA using Lipofectamine RNAiMAX (13778-150, Thermo Scientific, Waltham, MA, USA) according to the manufacturer’s protocol. siSIRT1 (pre-designed, 23411, Bioneer, Daejun, Korea); siMfn1 (pre-designed, 55669, Bioneer, Daejun, Korea) or control RNA (siCtrl; CCUACGCCACCAAUUUCGU (dTdT)) (Bioneer, Daejun, Korea) were used.

Techniques: Transfection, Incubation, Microscopy, Staining, Flow Cytometry, Produced, Phospho-proteomics