Fig 1: Chronic administration of SAHA decreases HDAC4 levels and reduces aggregate load – 1st trial.HDAC4 protein levels are reduced in the cortex and brain stem, but not in the hippocampus of WT and R6/2 mice treated with SAHA. Representative western immunoblots of 20 µg of cortical, brain stem and hippocampal homogenates from 9-week-old WT (A) and R6/2 (B) mice treated with SAHA or vehicle, immunoprobed with the Sigma (Fig. 1A, 1B - brain stem) or the Santa Cruz (Fig. 1A, 1B – cortex and hippocampus) anti-HDAC4 antibodies are shown. HDAC4 protein levels were normalized to alpha-tubulin and quantified using densitometry. Representative graphs illustrate relative HDAC4 protein levels as a percentage of the respective vehicle group. Error bars represent S.E.M. (n = 4). The seprion-ligand ELISA was used to quantify aggregate loads, which were significantly reduced in the cortex and brain stem but not in the hippocampus of the R6/2 SAHA treated mice (C). Error bars are S.E.M. (WT vehicle: n = 6; WT SAHA: n = 6; R6/2 vehicle: n = 6; R6/2 SAHA n = 5).
Fig 2: Chronic administration of SAHA decreases HDAC4 protein but not mRNA levels – 2nd trial.HDAC4 protein levels are reduced in the cortex and brain stem, but not in the hippocampus of WT and R6/2 mice treated with SAHA. Representative western immunoblots of 20 µg of cortical, brain stem and hippocampal homogenates from 9-week-old WT (A) and R6/2 (B) mice treated with SAHA or vehicle, immunoprobed with the Santa Cruz anti-HDAC4 antibody are shown. HDAC4 protein levels were normalized to alpha-tubulin and quantified using densitometry. Representative graphs illustrate relative HDAC4 protein levels as a percentage of the respective vehicle group. Error bars represent S.E.M. (n = 4). C. Hdac4 transcript levels were unaffected in the cortex and brain stem of the WT and R6/2 SAHA treated mice. Hdac4 mRNA expression levels are shown as a relative expression ratio to the geometric mean of three housekeeping genes in the cortex and brain stem of 9-week-old WT vehicle (red bars), WT SAHA (blue bars), R6/2 vehicle (yellow bars) and R6/2 SAHA (green bars) mice. Error bars are S.E.M (n = 6). NS = not specific.
Supplier Page from MilliporeSigma for Anti-Histone Deacetylase 4 (HDAC4) (DM-15) antibody produced in rabbit