Journal: Cell reports

Article Title: ATAC and SAGA co-activator complexes utilize co-translational assembly, but their cellular localization properties and functions are distinct

doi: 10.1016/j.celrep.2023.113099

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: rabbit polyclonal anti-acetylated lysine , Cell Signaling Technology , 9441; RRID: AB_331805.

Techniques: Recombinant, Plasmid Preparation, RNA Extraction, Western Blot, Mass Spectrometry, Expressing, Software

Journal: Cell & Bioscience

Article Title: NADH elevation during chronic hypoxia leads to VHL-mediated HIF-1α degradation via SIRT1 inhibition

doi: 10.1186/s13578-023-01130-3

Figure Lengend Snippet: SIRT1 deacetylates Lys709 of HIF-1α to facilitate protein stabilization, even under conditions of protein dehydroxylation. (A) Overexpression of p300 regulated HIF-1a stability. HIF-1α protein levels were determined under hypoxic exposure for 9 h after overexpression of p300. (B) Deacetylation of HIF-1α by SIRT1. 293T cells were transiently transfected with Flag-tagged HIF-1α and HA-tagged p300 together with wt-SIRT1 (WT) or SIRT1/H363Y (DN). HIF-1α acetylation and HIF-1α-SIRT1 interaction in immunoprecipitated HIF-1α were assessed by probing with antibodies to acetylated lysine (AcK), and SIRT1, respectively. (C) 293T cells were transiently cotransfected with 2 µg of each of Flag-tagged HIF-1α , K709R or empty vector, and 2 µg of each of HA-tagged p300 and/or Myc-tagged SIRT1. (D) The stability of K709Q was compared to that of K709R and wt-HIF-1α in SIRT1-depleted HeLa cell clone either treated or not with MG132. (E) Acetylation and ubiquitination of immunoprecipitated WT- HIF-1α and K709R were assessed in in SIRT1-depleted HeLa cell clone

Article Snippet: Immunoblotting using an anti-acetyl-lysine (AcK) antibody (Cat. no. 9441, Cell Signaling, MA) that specifically recognizes acetylated lysines was performed for detection of acetylated HIF-1α.

Techniques: Over Expression, Transfection, Immunoprecipitation, Plasmid Preparation