Journal: Current Protocols

Article Title: Epigenetic Histone Deacetylases Activity Assay in the Brain and Peripheral Organ Tissues

doi: 10.1002/cpz1.70143

Figure Lengend Snippet: Biochemical basis of the HDAC activity assay in brain and other tissues. Figure created using BioRender.

Article Snippet: HDAC activity is measured in nuclear lysates using the fluorometric HDAC substrate Boc‐Lys(Ac)‐AMC/Boc‐Lys‐AMC assay (I‐1875 and I‐1880, Bachem), as reported previously (Nian et al., ; Rajendran et al., , ).

Techniques: HDAC Activity Assay

Journal: Current Protocols

Article Title: Epigenetic Histone Deacetylases Activity Assay in the Brain and Peripheral Organ Tissues

doi: 10.1002/cpz1.70143

Figure Lengend Snippet: Illustration of critical steps involved in measuring HDAC activity in the mice tissues. Figure created using BioRender.

Article Snippet: HDAC activity is measured in nuclear lysates using the fluorometric HDAC substrate Boc‐Lys(Ac)‐AMC/Boc‐Lys‐AMC assay (I‐1875 and I‐1880, Bachem), as reported previously (Nian et al., ; Rajendran et al., , ).

Techniques: Activity Assay

Journal: Current Protocols

Article Title: Epigenetic Histone Deacetylases Activity Assay in the Brain and Peripheral Organ Tissues

doi: 10.1002/cpz1.70143

Figure Lengend Snippet: Assay layouts for protein estimation and HDAC activity. ( A ) Representative sample layout of the 96‐well microplate for the BCA assay. ( B ) Representative sample layout of the 96‐well microplate for the HDAC activity assay. ( C ) A typical standard curve generated using the Spectra MAX Gemini XS Plate Reader is shown with standard concentrations of 50, 250, 500, 1000, 1500, and 2000 µg/ml at an excitation wavelength of 360 nm and emission wavelength of 460 nm. Panels A and B were created using BioRender.

Article Snippet: HDAC activity is measured in nuclear lysates using the fluorometric HDAC substrate Boc‐Lys(Ac)‐AMC/Boc‐Lys‐AMC assay (I‐1875 and I‐1880, Bachem), as reported previously (Nian et al., ; Rajendran et al., , ).

Techniques: Activity Assay, BIA-KA, HDAC Activity Assay, Generated

Journal: Current Protocols

Article Title: Epigenetic Histone Deacetylases Activity Assay in the Brain and Peripheral Organ Tissues

doi: 10.1002/cpz1.70143

Figure Lengend Snippet: Quantification of HDAC activity in brain and other tissues. ( A ) Basal HDAC activity in sham brain tissues 7 days after TBI. The spleen is shown to have the highest normalized HDAC activity. ( B ) Percentage HDAC activity normalized to hippocampus levels. ( C and D ) Inhibition of HDAC activity by HDAC inhibitors 24 hr after TBI in the hippocampus of male mice is shown as ( C ) normalized HDAC activity and ( D ) percentage change in HDAC activity. ( E ) Normalized HDAC activity in the cortex and hippocampus of male and female mice 24 hr after TBI. Data were normalized to samples with trichostatin A (TSA) and are presented as normalized HDAC activity. TBI, traumatic brain injury; ROMI, romidepsin; SAHA, vorinostat; SB, sodium butyrate; FEU, fluorescence emission unit. Data represent mean ± S.E.M. ( n = 6 to 8 for each group). In subpanels C and D, * p < .05 versus sham; # p < .05 versus TBI. In subpanel E, * p < .05 versus sham (same sex); # p < .05 versus TBI (across sex); & p < .05 versus sham (across sex). Data were analyzed using one‐way ANOVA followed by Tukey's post hoc test.

Article Snippet: HDAC activity is measured in nuclear lysates using the fluorometric HDAC substrate Boc‐Lys(Ac)‐AMC/Boc‐Lys‐AMC assay (I‐1875 and I‐1880, Bachem), as reported previously (Nian et al., ; Rajendran et al., , ).

Techniques: Activity Assay, Inhibition, Fluorescence

Journal: Journal of Psychiatry & Neuroscience : JPN

Article Title: Permissive epigenetic regulatory mechanisms at the histone level are enhanced in postmortem dorsolateral prefrontal cortex of individuals with schizophrenia

doi: 10.1503/jpn.230054

Figure Lengend Snippet: (A) Histone deacetylase (HDAC) activity in the dorsolateral prefrontal cortex of individuals with schizophrenia (SZ) and matched controls (CTL) adjusted to Michaelis–Menten nonlinear regression model. Values are expressed as converted substrate in nM every minute per μg protein. HDAC activity parameters for (B) maximal velocity (V max ) and (C) Michaelis constant (K M ). Data correspond to 18 individuals with schizophrenia (10 antipsychotic [AP]-free, 8 antipsychotic-treated) and 18 matched controls.* p < 0.05 v. matched controls (2-tailed paired t test). ** p < 0.01 v. matched controls (2-tailed paired t test).

Article Snippet: HDAC activity was measured in the nucleosolic fraction (S2) using the fluorometric HDAC substrate Boc-Lys(Ac)-AMC/Boc-Lys-AMC (I-1875 and I-1880, respectively; Bachem), as reported previously with minor modifications.

Techniques: Histone Deacetylase Assay, Activity Assay

Journal: Journal of Psychiatry & Neuroscience : JPN

Article Title: Permissive epigenetic regulatory mechanisms at the histone level are enhanced in postmortem dorsolateral prefrontal cortex of individuals with schizophrenia

doi: 10.1503/jpn.230054

Figure Lengend Snippet: Protein expression levels of (A) histone deacetylase 1 (HDAC1), (B) HDAC2, and (C) HDAC4 in the dorsolateral prefrontal cortex of individuals with schizophrenia (SZ) and matched controls (CTL). The data shown are mean values of 3–4 independent experiments and are expressed as corrected immunodensity values normalized for β-actin as well as to a standard sample (STD) loaded in the same gel. Representative images of studied HDAC immunodensity in antipsychotic (AP)-free and antipsychotic-treated individuals and in matched controls are shown. Data correspond to 21 individuals with schizophrenia (11 antipsychotic-free, 10 antipsychotic-treated) and 21 matched controls. * p < 0.05 v. matched controls (2-tailed paired t test).

Article Snippet: HDAC activity was measured in the nucleosolic fraction (S2) using the fluorometric HDAC substrate Boc-Lys(Ac)-AMC/Boc-Lys-AMC (I-1875 and I-1880, respectively; Bachem), as reported previously with minor modifications.

Techniques: Expressing, Histone Deacetylase Assay

Journal: Journal of Psychiatry & Neuroscience : JPN

Article Title: Permissive epigenetic regulatory mechanisms at the histone level are enhanced in postmortem dorsolateral prefrontal cortex of individuals with schizophrenia

doi: 10.1503/jpn.230054

Figure Lengend Snippet: Pairwise associations between K M and V max coefficients estimated from global HDAC enzymatic activity assays and the Western blot targets

Article Snippet: HDAC activity was measured in the nucleosolic fraction (S2) using the fluorometric HDAC substrate Boc-Lys(Ac)-AMC/Boc-Lys-AMC (I-1875 and I-1880, respectively; Bachem), as reported previously with minor modifications.

Techniques: Activity Assay, Western Blot, Control