Journal: Frontiers in Medicine

Article Title: Investigating the causal relationship of lipid metabolism in polycystic ovary syndrome: a Mendelian randomization study on the regulatory role of 3-Hydroxybutyrate in gene expression

doi: 10.3389/fmed.2026.1747593

Figure Lengend Snippet: (A) 3-HB; (B) RGFP966. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: HDAC3-specific inhibitor RGFP966 (MCE, Catalog No. HY-13909), 3-Hydroxybutyrate (MCE, Catalog No. HY-113378).

Techniques:

Journal: Leukemia

Article Title: GNAS knockout potentiates HDAC3 inhibition through viral mimicry-related interferon responses in lymphoma.

doi: 10.1038/s41375-024-02325-4

Figure Lengend Snippet: Fig. 4 GNAS KO synergizes with HDAC3 inhibition to enhance CD8+ T cell-induced cytotoxicity. A Schematic representation of the T cell co-culture cytotoxicity assay. B Quantification of percentage cell death in DLBCL cells. Error bars represent ± SEM (n = 2 independent experiments). *, p < 0.05; **, p < 0.01; ***, p < 0.001 (log2 transformed values or values were compared; one-way ANOVA with Tukey’s HSD test for multiple comparisons). C Flow cytometry-based determination of cell death. Single cells and non-debris were included for analysis of 5- (and 6)-Carboxyfluorescein diacetate succinimidyl ester (CFSE) and propidium iodide (PI) signals. CFSE positive and negative populations represent DLBCL cells and CD8+ T cells, respectively. PI positive and negative groups represent dead and live cells, respectively. The total number of single DLBCL cells acquired per sample was 10,000. The percentages shown are percentage cell death within the total DLBCL cell population per sample. PBMCs, peripheral blood mononuclear cells.

Article Snippet: The selective HDAC3 inhibitors RGFP966 and BRD3308 (Selleckchem), the pan-HDAC inhibitor romidepsin (Selleckchem), the cAMP analog 8-Br-cAMP (Tocris), the TBK1 degrader TBK1 PROTAC 3i (Tocris), the JAK1/2 inhibitor ruxolitinib (Selleckchem), and the DNA methyltransferase inhibitor 5-Aza2’-deoxycytidine (5-AZA-CdR; MedChemExpress) were reconstituted and stored according to the manufacturer’s instructions.

Techniques: Inhibition, Co-Culture Assay, Cytotoxicity Assay, Transformation Assay, Flow Cytometry

Journal: Leukemia

Article Title: GNAS knockout potentiates HDAC3 inhibition through viral mimicry-related interferon responses in lymphoma.

doi: 10.1038/s41375-024-02325-4

Figure Lengend Snippet: Fig. 5 GNAS KO plus RGFP966 is associated with dsRNA formation and TE expression for viral mimicry induction. A Representative images of SU-DHL-4 cells stained by Hoechst 33342 (for cell nuclei; blue) and J2 antibody (for dsRNAs; green) from immunofluorescence analysis. Control LacZ cells omitting primary J2 antibody and treated with 300 nM 5-AZA-CdR were the negative and positive control, respectively. Scale bar, 25 µm. B Quantification of results in A. **p < 0.01; ****p < 0.0001 (Kruskal–Wallis test with Wilcoxon rank sum test for pairwise comparisons). C Volcano plots showing normalized expression dynamics of differentially expressed TEs (FDR < 0.05 and |Log2 fold change | > 0) identified from TEtranscripts analysis of cell-line bulk RNA-seq data. Upregulated and downregulated TEs are highlighted in red and blue, respectively. The horizontal and vertical dotted lines indicate FDR = 0.05 and Log2 fold change = 0, respectively. D Normalized TE expression under each comparison by a ratio of upregulated versus downregulated TEs shown in C. The dotted line represents normalized TE expression = 1 (equal numbers of upregulated and downregulated TEs).

Article Snippet: The selective HDAC3 inhibitors RGFP966 and BRD3308 (Selleckchem), the pan-HDAC inhibitor romidepsin (Selleckchem), the cAMP analog 8-Br-cAMP (Tocris), the TBK1 degrader TBK1 PROTAC 3i (Tocris), the JAK1/2 inhibitor ruxolitinib (Selleckchem), and the DNA methyltransferase inhibitor 5-Aza2’-deoxycytidine (5-AZA-CdR; MedChemExpress) were reconstituted and stored according to the manufacturer’s instructions.

Techniques: Expressing, Staining, Control, Positive Control, RNA Sequencing, Comparison

Journal: Molecular Neurobiology

Article Title: HDAC3 Inhibition Stimulates Myelination in a CMT1A Mouse Model

doi: 10.1007/s12035-022-02782-x

Figure Lengend Snippet: Inhibiting HDAC3 enzyme activity increases AKT-ERK1/2 signaling and myelin basic protein expression. a Schematic overview of the treatment scheme for the C3 and wild-type mice (Wt) with RGFP966. Treatment consisted of 1 week of daily subcutaneous injections, followed by I.P. injections given every second day for a total of 2 weeks (7 I.P. injections in total). Treatment is represented by a full green line followed by 7 green dashes representing the periodic distribution of treatment injections. Phenotyping is represented as a solid blue line, representing a week-long examination of the animals’ motor phenotype starting from postnatal day 30, 2 days after the treatment has been completed. This was followed by electrophysiological examinations and, lastly, by sacrificing of the animals for histopathological examinations. b Western blot of sciatic nerve cytoplasmic and nuclear-enriched fractions from a Wt control (Wt ctrl) and a Wt treated mouse with 10 mg/kg RGFP966. Acetylated histone H3 (Ac-H3) and histone H4 are shown in the nuclear-enriched fraction. Histone H4 was used as loading control for the nuclear fraction and for quantifications. MBP, p-ERK1/2, ERK1/2, p-AKT, and AKT are shown in the cytoplasmic-enriched fractions. A total protein stain was used as loading control. c Quantification of Western blots shown in b ( n = 4 mice for each group). All quantifications are expressed as fold change compared to Wt ctrl. d Schematic overview showing PI3K/AKT and MAPK/ERK1/2 signaling pathways during the initiation and active myelination, based on and reviewed in . Image was created with http://www.BioRender.com . HDAC3 dotted lines are based on protein expression shown in b-c , as well as from He et al.

Article Snippet: For treatment, the selective HDAC3 inhibitor (HDAC3i) RGFP966 (99.5% purity; Asclepia, Destelbergen, Belgium) was used as reported before [ , ].

Techniques: Activity Assay, Expressing, Western Blot, Control, Staining, Protein-Protein interactions

Journal: Molecular Neurobiology

Article Title: HDAC3 Inhibition Stimulates Myelination in a CMT1A Mouse Model

doi: 10.1007/s12035-022-02782-x

Figure Lengend Snippet: Inhibiting HDAC3 enzyme activity increases AKT-ERK1/2 signaling and myelin proteins during C3 postnatal development. a Western blots of p-AKT, AKT, p-ERK, and ERK1/2 in addition to the myelin proteins MBP, P0, and PMP22, in Wt ctrl, C3 ctrl and C3 5 mg/kg (left), and Wt ctrl, C3 ctrl, and 10 mg/kg RGFP966 treated mice (right). b – f Quantifications of the Western blots. Additional blots used in quantifications relating to C3 5 mg/kg treated mice as well as C3 10 mg/kg treated mice are included in Additional file 2, Fig. . All myelin proteins were normalized to total protein stain, which is located below each Western blot. For b and c , Wt ctrl n = 11 mice, C3 ctrl n = 12 mice, C3 5 mg/kg n = 8 mice, C3 10 mg/kg n = 4 mice. For d , Wt ctrl n = 11 mice, C3 ctrl n = 12 mice, C3 5 mg/kg n = 8 mice, C3 10 mg/kg n = 4 mice. For e , Wt ctrl n = 15 mice, C3 ctrl n = 16 mice, C3 5 mg/kg n = 8 mice, C3 10 mg/kg n = 8 mice. For f , C3 ctrl n = 12 mice, C3 5 mg/kg n = 8 mice, C3 10 mg/kg n = 4 mice. Statistical significance was evaluated in b – e with a two-way ANOVA with a Tukey’s multiple comparison’s test (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). Statistical significance was evaluated in f with one-way ANOVA and Tukey’s multiple comparison’s test (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). In b – d , data are expressed as fold change from the mean of the Wt ctrl group ± SD. In f , data are expressed as fold change from the mean of the C3 ctrl group ± SD, as normalization to PMP22 expression in the Wt ctrl was not possible

Article Snippet: For treatment, the selective HDAC3 inhibitor (HDAC3i) RGFP966 (99.5% purity; Asclepia, Destelbergen, Belgium) was used as reported before [ , ].

Techniques: Activity Assay, Western Blot, Staining, Expressing

Journal: Molecular Neurobiology

Article Title: HDAC3 Inhibition Stimulates Myelination in a CMT1A Mouse Model

doi: 10.1007/s12035-022-02782-x

Figure Lengend Snippet: HDAC3 inhibition improves NCVs and CMAP latencies and amplitude recordings in C3 mice. a Representative tracings of electrophysiological recordings for the C3 ctrl and for the C3 mice treated with 5 mg/kg and 10 mg/kg of RGFP966. Arrowheads indicate the point at which the potential starts. All electrophysiological tests were conducted during phenotyping between postnatal days 35 and 36. b Quantification of the nerve conduction velocity (NCV) recordings from the sciatic nerve. c Quantification of CMAP latencies. d Quantifications of the CMAP amplitudes. In b – c , Wt ctrl values are indicated by a black dotted line in each graph. Statistical significance was determined in panel b with a Kruskal–Wallis test with Dunn’s multiple comparison’s test, in panels c and d with a one-way ANOVA with a Tukey’s multiple comparison’s test (* p < 0.05 and ** p < 0.01). Mice used in b – d per group: C3 ctrl = 33, C3 5 mg/kg = 23, and C3 10 mg/kg = 8. Data are expressed in b – d as mean ± SD

Article Snippet: For treatment, the selective HDAC3 inhibitor (HDAC3i) RGFP966 (99.5% purity; Asclepia, Destelbergen, Belgium) was used as reported before [ , ].

Techniques: Inhibition

Journal: Molecular Neurobiology

Article Title: HDAC3 Inhibition Stimulates Myelination in a CMT1A Mouse Model

doi: 10.1007/s12035-022-02782-x

Figure Lengend Snippet: HDAC3 inhibition increases myelination and axon caliber size in C3 mice. a Electron microscopy images of the sciatic nerve sections from the Wt ctrl, C3 ctrl, C3 5 mg/kg, and C3 10 mg/kg groups. Scale bar: 10 μm. Schwann cell dysmyelination is indicated by red arrowheads, hypermyelination by red asterisks and hypomyelination by black arrows. Images were taken at the central region of the sciatic nerve. b Analysis of g -ratios. C3 ctrl mice (red, n = 4 mice) compared with Wt ctrl mice (black, n = 4 mice). C3 5 mg/kg HDAC3i treated mice (green, n = 2) compared to C3 ctrl (second panel) and C3 10 mg/kg RGFP966 treated mice (blue, n = 4) compared to the C3 ctrl mice (third panel in from the left). Wt ctrl mice compared to C3 10 mg/kg HDAC3i treated mice (fourth panel to the right). A range of 150–200 fibers per animal were measured at 800 × magnification. c Histogram of the relative frequency distribution of the axon diameters. Kolmogorov–Smirnov, two-tailed statistical analysis was used to analyze the difference between the cumulative distribution curves of each sample. Wt ctrl vs C3 ctrl and C3 ctrl vs C3 10 mg/kg were reported in the figure. For C3 ctrl vs C3 5 mg/kg, p = 0.0031 and D = 0.1191. For Wt ctrl vs C3 10 mg/kg, p = < 0.0001 and D = 0.1746. For Wt ctrl vs C3 5 mg/kg, p = < 0.0001 and D = 0.2124. For C3 5 mg/kg vs C3 10 mg/kg, p = 0.0119, and D = 0.1056. d Quantifications of myelinated axons, e unmyelinated axons, and f total axons per 100 μm 2 per sciatic nerve section. Number of sciatic nerve sections analyzed d – f were as follows: Wt ctrl n = 7, C3 ctrl n = 8, and C3 10 mg/kg n = 11. Statistical significance was evaluated in d with a one-way ANOVA, Dunn’s multiple comparison’s test (* p < 0.05, **** p < 0.0001). Statistical significance was evaluated in e and f with a Kruskal–Wallis test, with Dunn’s multiple comparison test (* p < 0.05) to compare groups. Total mice used in b – f per group: Wt ctrl = 4, C3 ctrl = 4, C3 5 mg/kg = 2, and C3 10 mg/kg = 4. Total number of nerve fibers analyzed in b and c per group: Wt ctrl = 552, C3 ctrl = 652, C3 5 mg/kg = 350, C3 10 mg/kg = 698

Article Snippet: For treatment, the selective HDAC3 inhibitor (HDAC3i) RGFP966 (99.5% purity; Asclepia, Destelbergen, Belgium) was used as reported before [ , ].

Techniques: Inhibition, Electron Microscopy, Two Tailed Test, Comparison

Journal: Molecular Neurobiology

Article Title: HDAC3 Inhibition Stimulates Myelination in a CMT1A Mouse Model

doi: 10.1007/s12035-022-02782-x

Figure Lengend Snippet: High-dose HDAC3 inhibition causes a decline in the motor performance in Wt and C3 mice. a Effect of RGFP966 on overall grip strength was assessed on the Wt mice at postnatal day 30. b At the same time-point, the effect of RGFP966 on the motor performance was assessed using rotarod analysis in the Wt mice. For the C3 mouse model (with the Wt ctrl group included as reference), overal grip strength c , and rotarod performance d , was assessed. Statistical significance was evaluated in b and d with a Kruskal–Wallis test with Dunn’s multiple comparison’s test (** p < 0.01, **** p < 0.0001). Statistical significance was evaluated in a and c with a one-way ANOVA with a Tukey’s multiple comparison’s test (* p < 0.05, ** p < 0.01, **** p < 0.0001). Mice used in the different groups in panels a - d per group: Wt ctrl, n = 32; Wt 5 mg/kg, n = 16; Wt 10 mg/kg, n = 5; C3 ctrl, n = 33; C3 5 mg/kg, n = 23; and C3 10 mg/kg, n = 8

Article Snippet: For treatment, the selective HDAC3 inhibitor (HDAC3i) RGFP966 (99.5% purity; Asclepia, Destelbergen, Belgium) was used as reported before [ , ].

Techniques: Inhibition

Journal: Molecular Neurobiology

Article Title: HDAC3 Inhibition Stimulates Myelination in a CMT1A Mouse Model

doi: 10.1007/s12035-022-02782-x

Figure Lengend Snippet: HDAC3 inhibition increases the presence of endoneural macrophages in C3 but not in wild-type mice. a Immunohistochemical staining in the brachial plexus nerves of CD34+ cells (fibroblasts) in red, F4/80+ cells (endoneural macrophage) in green, and the nucleus of cells are stained with DAPI in blue. Scale bars: 100 μm. A white dashed oval encompasses the region of the sections analyzed. To the right of each image is a 400% zoomed focus of areas to highlight F4/80+ cells (white arrowheads) and the interaction between CD43+ and F4/80+ cells (white asterisk). Scale bar: 20 µm. b Macrophage presence was quantified by the number of F4/80+ cells per slice, and macrophage activation was quantified by the overlap between CD34+ and F4/80+ cells per slice. c Western blot analysis of sciatic nerve CSF-1-R protein expression in C3 ctrl and 10 mg/kg treated mice, with quantification of the Western blot below. Western blot quantification is expressed as fold change from C3 ctrl. For b , data are represented as the mean of 3–4 slices for each mouse ± SEM. The number of mice analyzed was as follows: Wt ctrl n = 6, Wt 10 mg/kg n = 4, C3 ctrl n = 5, and C3 10 mg/kg n = 6. For statistical significance in b , a two-way ANOVA with a Tukey’s multiple comparison’s test (* p < 0.05, **** p < 0.0001) was used to compare groups. In c , a student’s t-test was used to determine statistical significance (* p < 0.05)

Article Snippet: For treatment, the selective HDAC3 inhibitor (HDAC3i) RGFP966 (99.5% purity; Asclepia, Destelbergen, Belgium) was used as reported before [ , ].

Techniques: Inhibition, Immunohistochemical staining, Staining, Activation Assay, Western Blot, Expressing

Journal: Scientific Reports

Article Title: A selective inhibitor of histone deacetylase 3 prevents cognitive deficits and suppresses striatal CAG repeat expansions in Huntington’s disease mice

doi: 10.1038/s41598-017-05125-2

Figure Lengend Snippet: Systemic RGFP966 treatment efficiently inhibits hippocampal and striatal HDAC activity. (a) Schematic of chronic treatment and experimental plan. ARTP, accelerating rotarod task procedure; OLT, object location task; NORT, novel object recognition task. ( b) Body weight measurements. Data represent the mean ± SEM (n = 6–10). ( c) Representative immunoblots showing histone H3 acetylation levels at position lysine 9 (AcH3K9) normalized to total histone H3 and (d) HDAC3 levels normalized to α-tubulin in striatal and hippocampal extracts from vehicle and RGFP966 chronically treated wild type Hdh Q7/Q7 (WT) and mutant Hdh Q7/Q111 (KI) mice. The blots in (c) and (d) are cropped; full-length images are provided in Supplementary Figure . * p < 0.05; ** p < 0.01; *** p < 0.001 compared to vehicle-treated mice by two-way ANOVA with Bonferroni post-hoc analysis. Data represent the mean ± SEM (n = 4–7 per group).

Article Snippet: The HDAC3 inhibitor RGFP966 was generously provided by BioMarin Pharmaceutical Inc. (San Rafael, California, USA).

Techniques: Activity Assay, Western Blot, Mutagenesis

Journal: Scientific Reports

Article Title: A selective inhibitor of histone deacetylase 3 prevents cognitive deficits and suppresses striatal CAG repeat expansions in Huntington’s disease mice

doi: 10.1038/s41598-017-05125-2

Figure Lengend Snippet: RGFP966 treatment prevents impairment of long-term memory in HD mice. Behavior analysis in vehicle and RGFP966 chronically treated Hdh Q7/Q7 (WT) and Hdh Q7/Q111 (KI) mice. Performance in (a) spatial (OLT) and (b) recognition (NORT) memory tests. Exploration time during training and testing session (left and center panels). Dashed line shows chance level for exploration. *** p < 0.001 compared to the percentage of time exploring the familiar object by two-way ANOVA with Bonferroni post-hoc analysis. Discrimination index (right panel). * p < 0.05, *** p < 0.001 compared to vehicle-treated KI mice; # p < 0.05 compared to vehicle-treated WT mice by two-way ANOVA with Bonferroni post-hoc analysis. Data represent the mean ± SEM (n = 5–9 animals per group).

Article Snippet: The HDAC3 inhibitor RGFP966 was generously provided by BioMarin Pharmaceutical Inc. (San Rafael, California, USA).

Techniques:

Journal: Scientific Reports

Article Title: A selective inhibitor of histone deacetylase 3 prevents cognitive deficits and suppresses striatal CAG repeat expansions in Huntington’s disease mice

doi: 10.1038/s41598-017-05125-2

Figure Lengend Snippet: RGFP966 treatment restores gene expression in the hippocampus of HD mice. (a) Schematic time line of acute treatment. ( b) Quantitative RT-PCR analysis of memory-related genes in the hippocampus of vehicle and RGFP966 acutely treated Hdh Q7/Q7 (WT) and Hdh Q7/Q111 (KI) mice. Histogram represents relative mRNA abundance of Arc, Nr4a2, Egr1 and c-Fos . Levels of mRNA were normalized to 18S and Actinβ. * p < 0.05 compared to vehicle-treated WT mice by two-way ANOVA with Bonferroni post-hoc analysis. (c) Representative images (high magnification) showing Arc immunostaining in the hippocampal dentate gyrus of acutely treated Hdh Q7/Q7 (WT) and Hdh Q7/Q111 (KI) mice. Histogram shows quantification of the average number of Arc-positive neurons in the dentate gyrus. * p < 0.05 compared to vehicle-treated WT mice by two-way ANOVA with Bonferroni post-hoc analysis. (d) Representative images with magnification insets of the hippocampal CA1 region showing Egr1 immunostaining in acutely treated Hdh Q7/Q7 (WT) and Hdh Q7/Q111 (KI) mice. Histogram shows quantification of the average intensity of Egr1 immunoreactivity in the CA1 or the CA3 region. Data represent the mean ± SEM (n = 5–8 animals per group).

Article Snippet: The HDAC3 inhibitor RGFP966 was generously provided by BioMarin Pharmaceutical Inc. (San Rafael, California, USA).

Techniques: Expressing, Quantitative RT-PCR, Immunostaining

Journal: Scientific Reports

Article Title: A selective inhibitor of histone deacetylase 3 prevents cognitive deficits and suppresses striatal CAG repeat expansions in Huntington’s disease mice

doi: 10.1038/s41598-017-05125-2

Figure Lengend Snippet: RGFP966 treatment induces Arc but not Egr1 or c-Fos protein levels in primary hippocampal cultures. (a ) Representative immunoblots showing histone H3 acetylation levels at position lysine 9 (AcH3K9) with actin as loading control and (b) Arc, Egr1 and c-Fos protein levels normalized to actin levels in DMSO and RGFP966 treated primary hippocampal cultures obtained from Hdh Q7/Q7 (WT) and Hdh Q7/Q111 (KI) embryos. The blots in (a and b) are cropped; full-length images are provided in Supplementary Figure . ** p < 0.01 compared to DMSO-treated cultures by two-way ANOVA with Bonferroni post-hoc analysis. Data represent the mean ± SEM (n = 4–5 cultures per group).

Article Snippet: The HDAC3 inhibitor RGFP966 was generously provided by BioMarin Pharmaceutical Inc. (San Rafael, California, USA).

Techniques: Western Blot

Journal: Scientific Reports

Article Title: A selective inhibitor of histone deacetylase 3 prevents cognitive deficits and suppresses striatal CAG repeat expansions in Huntington’s disease mice

doi: 10.1038/s41598-017-05125-2

Figure Lengend Snippet: RGFP966 treatment prevents impairment of motor learning in HD mice. Behavior analysis in vehicle and RGFP966 chronically treated Hdh Q7/Q7 (WT) and Hdh Q7/Q111 (KI) mice. (a) Latency to fall in the accelerating rotarod task. ** p < 0.01 compared to vehicle-treated WT mice by two-way ANOVA with repeated measurements. (b) Spontaneous locomotor activity measured by total distance traveled during two consecutive days (15 min/day). Data represent the mean ± SEM (n = 5–9 animals per group).

Article Snippet: The HDAC3 inhibitor RGFP966 was generously provided by BioMarin Pharmaceutical Inc. (San Rafael, California, USA).

Techniques: Activity Assay

Journal: Scientific Reports

Article Title: A selective inhibitor of histone deacetylase 3 prevents cognitive deficits and suppresses striatal CAG repeat expansions in Huntington’s disease mice

doi: 10.1038/s41598-017-05125-2

Figure Lengend Snippet: RGFP966 suppresses striatal CAG repeat expansions. (a) Representative display of small-pool PCR of somatic repeat lengths from single Hdh Q7/Q111 (KI) animals, either chronically treated with vehicle or with RGFP966. Size markers correspond to CAG repeat number. The examples shown represent the positive signals from one analysis. Arrow indicates size of initial CAG tract. This image has been cropped; the original is provided in Supplementary Figure . ( b ) Summary of genetic changes. For each group, n = 3 mice, 62–65 independent events. Starting tract sizes, estimated from the most common allele size in cerebellum and stated as repeat units, were 110–119, 120–129 and 120–129 for vehicle-treated mice; and 120–129, 120–129 and 130–139 for RGFP966-treated mice. * p < 0.05 by Student’s t-test. (c) Histogram of expansion sizes from the alleles tested in b . Allele sizes were binned into 10-repeat size groups, e.g . falling between +1 to +10 repeats compared to the starting allele size. Starting tract lengths were deduced from the most common allele size in the cerebellum of the same animals.

Article Snippet: The HDAC3 inhibitor RGFP966 was generously provided by BioMarin Pharmaceutical Inc. (San Rafael, California, USA).

Techniques:

Journal: Scientific Reports

Article Title: A selective inhibitor of histone deacetylase 3 prevents cognitive deficits and suppresses striatal CAG repeat expansions in Huntington’s disease mice

doi: 10.1038/s41598-017-05125-2

Figure Lengend Snippet: RGFP966 treatment ameliorates striatal pathology in HD mice. (a) Representative immunoblots showing striatal protein levels of DARPP-32, PDE10A and A 2A R, with actin as loading control in vehicle and RGFP966 chronically treated Hdh Q7/Q7 (WT) and Hdh Q7/Q111 (KI) mice. * p < 0.05 compared to vehicle-treated KI mice; # p < 0.05; ### p < 0.001 compared to vehicle treated WT mice by two-way ANOVA with Bonferroni post-hoc analysis. Data represent the mean ± SEM (n = 6–11 animals per group). (b) Representative immunoblots showing striatal protein levels of DARPP-32, PDE10A and A 2A R with actin as loading control in DMSO and RGFP966 treated primary striatal cultures obtained from Hdh Q7/Q7 (WT) and Hdh Q7/Q111 (KI) embryos. * p < 0.05 and *** p < 0.001 compared to DMSO-treated cultures by two-way ANOVA with Bonferroni post-hoc analysis. Data represent the mean ± SEM (n = 4–5 cultures per group). (c) Representative immunoblots showing oligomeric forms of mutant huntingtin (stacking gel), monomeric mutant huntingtin (resolving gel) and α-tubulin as loading control in vehicle and RGFP966 chronically treated Hdh Q7/Q111 (KI) mice. Samples from two different mice are shown in adjoining lanes. All blots have been cropped; the original images are shown in Supplementary Figure . * p < 0.05 by Student’s t-test. Data represent the mean ± SEM (n = 6–10 animals per group). (d) Quantitative RT-PCR analysis of Htt mRNA in the striatum of vehicle and RGFP966 acutely treated Hdh Q7/Q7 (WT) and Hdh Q7/Q111 (KI) mice. Levels of mRNA were normalized to 18S and Actinβ. Data represent the mean ± SEM (n = 6–7 animals per group).

Article Snippet: The HDAC3 inhibitor RGFP966 was generously provided by BioMarin Pharmaceutical Inc. (San Rafael, California, USA).

Techniques: Western Blot, Mutagenesis, Quantitative RT-PCR