Journal: PLoS ONE

Article Title: Class I histone deacetylase inhibitor MS-275 attenuates vasoconstriction and inflammation in angiotensin II-induced hypertension

doi: 10.1371/journal.pone.0213186

Figure Lengend Snippet: IC 50 [μM] values for TMP269, RGFP966, and TSA.

Article Snippet: To evaluate the HDAC enzyme inhibitory activity of MS-275 and RGFP966, we determined enzyme activities of HDAC1, HDAC2, HDAC3, and HDAC8 using enzyme assay kits (BPS Bioscience, San Diego, CA, USA) according to the manufacturer’s protocols.

Techniques:

Journal: PLoS ONE

Article Title: Class I histone deacetylase inhibitor MS-275 attenuates vasoconstriction and inflammation in angiotensin II-induced hypertension

doi: 10.1371/journal.pone.0213186

Figure Lengend Snippet: (A) After angiotensin II infusion (1.3 mg·kg -1 ·day -1 ) to mice for 1 week, we injected MS-275 or RGFP966 (both 3 mg·kg -1 ·day -1 ) daily to mice for additional 7 days. Systolic blood pressures were measured in awake mice. (B and C) Transcript levels of aortic AT1 and ACE1 were quantified using quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). GAPDH was used to normalize the values. Data are presented as the means ± SE (n = 8 per group). ***p < 0.001 versus sham group; # p < 0.05 and ### p < 0.001 versus angiotensin II group; NS, not significant.

Article Snippet: To evaluate the HDAC enzyme inhibitory activity of MS-275 and RGFP966, we determined enzyme activities of HDAC1, HDAC2, HDAC3, and HDAC8 using enzyme assay kits (BPS Bioscience, San Diego, CA, USA) according to the manufacturer’s protocols.

Techniques: Injection, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: Class I histone deacetylase inhibitor MS-275 attenuates vasoconstriction and inflammation in angiotensin II-induced hypertension

doi: 10.1371/journal.pone.0213186

Figure Lengend Snippet: (A) Representative images of H&E stained aortas from sham, angiotensin II group (Ang II), MS-275-treated angiotensin II group (Ang II + MS-275), and RGFP966-treated angiotensin II group (Ang II + RGFP966). Scale bar = 100 μm. (B) Arterial wall thickness was quantified. Data are means ± SE (n = 7/group). ***p < 0.001 versus sham group; ## p < 0.01 and ### p < 0.001 versus angiotensin II group. (C‒D) Masson’s trichrome and orcein staining of representative aorta sections. Scale bar = 100 μm. Collagen deposition in aorta is shown as blue staining. (E‒H) The transcript levels of cyclin D1, cyclin E1, E2F3, and GATA6 were quantified using quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). * p < 0.05 versus sham group; # p < 0.05 versus angiotensin II group; NS, not significant.

Article Snippet: To evaluate the HDAC enzyme inhibitory activity of MS-275 and RGFP966, we determined enzyme activities of HDAC1, HDAC2, HDAC3, and HDAC8 using enzyme assay kits (BPS Bioscience, San Diego, CA, USA) according to the manufacturer’s protocols.

Techniques: Staining, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: Class I histone deacetylase inhibitor MS-275 attenuates vasoconstriction and inflammation in angiotensin II-induced hypertension

doi: 10.1371/journal.pone.0213186

Figure Lengend Snippet: The transcript levels for arginase 1 (A), arginase 2 (B), GTPCH1 (C), PRMT1 (D), DDAH1 (E), and DDAH2 (F) were determined using quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) in aortas of sham and angiotensin II treated with vehicle, MS-275, or RGFP966. Data are presented as the means ± SE (n = 8 per group). *p < 0.05 and ***p < 0.001 versus sham group; # p < 0.05 versus angiotensin II group; NS, not significant.

Article Snippet: To evaluate the HDAC enzyme inhibitory activity of MS-275 and RGFP966, we determined enzyme activities of HDAC1, HDAC2, HDAC3, and HDAC8 using enzyme assay kits (BPS Bioscience, San Diego, CA, USA) according to the manufacturer’s protocols.

Techniques: Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

Journal: PLoS ONE

Article Title: Class I histone deacetylase inhibitor MS-275 attenuates vasoconstriction and inflammation in angiotensin II-induced hypertension

doi: 10.1371/journal.pone.0213186

Figure Lengend Snippet: The transcript levels of Nox1 (A), Nox2 (B), Nox4 (C), p22phox (D), p47phox (E), Cox-2 (F), and SOD3 (G) were determined using qRT-PCR in aortas of sham and Ang II-induced mice treated with vehicle, MS-275, or RGFP966. Results are ± SE (n = 8 per group). * p < 0.05 and ** p < 0.01 versus sham group; NS, not significant.

Article Snippet: To evaluate the HDAC enzyme inhibitory activity of MS-275 and RGFP966, we determined enzyme activities of HDAC1, HDAC2, HDAC3, and HDAC8 using enzyme assay kits (BPS Bioscience, San Diego, CA, USA) according to the manufacturer’s protocols.

Techniques: Quantitative RT-PCR

Journal: PLoS ONE

Article Title: Class I histone deacetylase inhibitor MS-275 attenuates vasoconstriction and inflammation in angiotensin II-induced hypertension

doi: 10.1371/journal.pone.0213186

Figure Lengend Snippet: The transcript levels of iNOS (A), TNF-α (B), IL-1β (C), MCP-1 (D), VCAM-1 (E), and ICAM-1 (F) were determined using quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) in aortas of sham and Ang II-induced mice treated with vehicle, MS-275, or RGFP966. Results are means ± SE (n = 8 per group). * p < 0.05, ** p < 0.01, and *** p < 0.001 versus sham group; # p < 0.05 and ## p < 0.01 versus angiotensin II group; NS, not significant. (G) Representative aortic images for macrophage infiltration are shown. Scale bar = 50 μm.

Article Snippet: To evaluate the HDAC enzyme inhibitory activity of MS-275 and RGFP966, we determined enzyme activities of HDAC1, HDAC2, HDAC3, and HDAC8 using enzyme assay kits (BPS Bioscience, San Diego, CA, USA) according to the manufacturer’s protocols.

Techniques: Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: HDAC3 inhibition ameliorates spinal cord injury by immunomodulation

doi: 10.1038/s41598-017-08535-4

Figure Lengend Snippet: HDAC3 inhibition changes functional state of innate immune cells and leads to global inflammatory suppression in SCI. ( a ) Diagram of divergent macrophage responses and hypothesized role of HDAC3. M0: homeostatic macrophages. ( b ) Experimental scheme of HDAC3 inhibitor treatment after SCI. Three i.p. injections of RGFP966 were delivered. ( c , d ) Representative IHC images showing increased AcH3 levels at the injury site with RPGF966 treatment at 3 dpi after T8 dorsal column transection (SCI-T). Enlarged images in the boxed area are shown in ( d ). ( e , f ) Representative immunofluorescent images and quantification of the numbers of CD16/32 immune cells (green), CD206 immune cells (red), and the ratio of the two subpopulations at the injury site at 10 dpi after contusion injury (SCI-C). DAPI was used for nuclear counterstaining. Arrow: example of double positive cell (CD16/32+ CD206+). n = 8 mice per group, with 2–4 representative images averaged for each animal. *p < 0.05, Unpaired Student’s t-test. ( g ) Relative cytokine/chemokine levels as determined by cytokine array using lysates of injured spinal cord from vehicle or RFGP966-treated animals at 10 dpi after contusion SCI. ***p < 0.001, two-way ANOVA with Bonferroni post hoc correction (n = 4 pairs of mice). Orientation: R: Rostral, C: Caudal, D: Dorsal, V: Ventral. Scale bar: 50 µm ( c ), 25 µm ( d ), and 100 µm ( e ).

Article Snippet: The DRG neurons were treated with increasing doses of RGFP966 for 48 hr, and fixed and immunostained for β-tubulin III (1:500, clone TUJ1, MAB1195, R&D systems), and counterstained with DAPI.

Techniques: Inhibition, Functional Assay

Journal: Scientific Reports

Article Title: HDAC3 inhibition ameliorates spinal cord injury by immunomodulation

doi: 10.1038/s41598-017-08535-4

Figure Lengend Snippet: HDAC3 inhibition improves functional recovery and enhances neuroprotective phenotypes after SCI. ( a ) Behavioral assays demonstrate improved and sustained functional recovery by both BMS and TMS scores up to 30 dpi after contusion injury. Blue line in the BMS score denotes weight-bearing threshold. Repeated measures, two-way ANOVA with Bonferroni post hoc correction. ### p < 0.001. n = 10 for vehicle and n = 6 for RGFP966 cohort. ( b ) Representative images and quantification show increased density of axon fibers, identified by NF-H immunolabeling (green), in lesion center at 10 dpi after contusion injury (SCI-C). Lesion border is indicated by dashed lines. DAPI was used for nuclear counterstaining. Enlarged images of boxed areas are shown below. n = 3 mice for vehicle and 4 mice for RGFP966 cohort, with 1–4 representative images averaged for each animal. **p < 0.01, unpaired Student’s t-test. ( c – e ) Representative images and quantification of SCI scar constituents at 10 dpi after contusion: CSPG (blue), GFAP (red), and Fibronectin (green). Lesion border is indicated by dashed lines. The number of animals quantified is shown inside the bar graph, with 2–5 representative image averaged for each animal. Quantification showed a decrease in CSPG with RGFP966 treatment, p = 0.028, Mann Whitney test, but no significant change in GFAP and a trend of decrease in Fibronectin (unpaired Student’s t test). The astroglial scar size was also not significantly changed (unpaired Student’s t test). Scale bars: 100 µm ( b ), 50 µm (i and ii in b ), and 500 µm ( c ).

Article Snippet: The DRG neurons were treated with increasing doses of RGFP966 for 48 hr, and fixed and immunostained for β-tubulin III (1:500, clone TUJ1, MAB1195, R&D systems), and counterstained with DAPI.

Techniques: Inhibition, Functional Assay, Immunolabeling, MANN-WHITNEY

Journal: Scientific Reports

Article Title: HDAC3 inhibition ameliorates spinal cord injury by immunomodulation

doi: 10.1038/s41598-017-08535-4

Figure Lengend Snippet: HDAC3 inhibition does not affect axon growth potential. ( a ) Dose-response neurite outgrowth assays show that RGFP966 treatment of cultured DRG neurons for 48 hr does not increase average length of longest axon. p = 0.073, Kruskal-Walis test. ( b , c ) Schematic diagram of in vivo RGFP966 treatment followed by DRG culture ( b ). No significant changes were observed for average axonal length between the two groups ( c ). unpaired Student’s t test. n.s. not statistically significant. The numbers of DRG neurons quantified are shown inside each bar graph.

Article Snippet: The DRG neurons were treated with increasing doses of RGFP966 for 48 hr, and fixed and immunostained for β-tubulin III (1:500, clone TUJ1, MAB1195, R&D systems), and counterstained with DAPI.

Techniques: Inhibition, Cell Culture, In Vivo

Journal: Scientific Reports

Article Title: HDAC3 inhibition ameliorates spinal cord injury by immunomodulation

doi: 10.1038/s41598-017-08535-4

Figure Lengend Snippet: HDAC3 activity contributes to inflammatory activation of microglia. ( a – c ) Experimental scheme, representative images of immunocytochemistry, and quantification show that LPS stimulation results in reduced AcH3 levels in primary microglia by 4 hr. Pre-exposure to RGFP966 reverses LPS-triggered AcH3 reduction. The numbers inside the bar graph indicate the numbers of cells quantified from nine photographs for each experimental condition. Mann Whitney test. **p = 0.0079, ***p < 0.001. ( d – f ) Experimental scheme, representative images of immunocytochemistry and quantification show that LPS stimulation decreases CD206+ population (green) and a trend of increase in iNOS+ population (red). RGFP966 pre-treatment reversed the LPS-induced bias, leading to increased CD206+ population and deceased iNOS+ population. DAPI for nuclear counterstaining. Arrow: examples of double positive (iNOS+ CD206+) cells. The numbers inside the bar graph indicate the numbers of images quantified. Mann Whitey test and unpaired Student’s t test, respectively. ( g – i ) Experimental scheme and quantification show HDAC3 knockdown by siRNA in primary microglia (unpaired Student’s t test). HDAC3 knockdown abolishes the polarized responses of microglia to LPS stimulation. The numbers inside the bar graph indicate the numbers of images quantified by Mann Whitney tests. Scale bar: 25 µm ( a ) and 50 µm ( e ).

Article Snippet: The DRG neurons were treated with increasing doses of RGFP966 for 48 hr, and fixed and immunostained for β-tubulin III (1:500, clone TUJ1, MAB1195, R&D systems), and counterstained with DAPI.

Techniques: Activity Assay, Activation Assay, Immunocytochemistry, MANN-WHITNEY, Knockdown

Journal: Microbiology Spectrum

Article Title: Histone deacetylase 2 and 3 of Sarcoptes scabiei : characterization of a potential drug target

doi: 10.1128/spectrum.00737-24

Figure Lengend Snippet: Survival curve of isolated scabies mites under different concentration of RGFP966 and Elevenostat.

Article Snippet: Elevenostat and RGFP966 (GlpBio, USA) were diluted to 10 µM, 1 µM, 100 nM, and 10 nM with deacetylation assays buffer for evaluating the inhibitory effects of Elevenostat and RGFP966 on rSsHDAC-2 and rSsHDAC-3, respectively.

Techniques: Isolation, Concentration Assay

Journal: Microbiology Spectrum

Article Title: Histone deacetylase 2 and 3 of Sarcoptes scabiei : characterization of a potential drug target

doi: 10.1128/spectrum.00737-24

Figure Lengend Snippet: Survival curves of Sarcoptes scabiei at different stages of development treated with 1 mg/mL RGFP966 and Elevenostat.

Article Snippet: Elevenostat and RGFP966 (GlpBio, USA) were diluted to 10 µM, 1 µM, 100 nM, and 10 nM with deacetylation assays buffer for evaluating the inhibitory effects of Elevenostat and RGFP966 on rSsHDAC-2 and rSsHDAC-3, respectively.

Techniques:

Journal: Microbiology Spectrum

Article Title: Histone deacetylase 2 and 3 of Sarcoptes scabiei : characterization of a potential drug target

doi: 10.1128/spectrum.00737-24

Figure Lengend Snippet: Ultrastructural changes of Sarcoptes scabies after 12 h treatment with RGFP966 and Elevenostat. ( A )10% DMSO; ( B )1 mg/mL RGFP966; ( C )1 mg/mL Elevenostat. Notes: N, nucleus; M, mitochondria; V, vacuole; LD, lipid droplet. Scale bars: A, B, and C = 2 µm; A, B, and C insets = 500 nm.

Article Snippet: Elevenostat and RGFP966 (GlpBio, USA) were diluted to 10 µM, 1 µM, 100 nM, and 10 nM with deacetylation assays buffer for evaluating the inhibitory effects of Elevenostat and RGFP966 on rSsHDAC-2 and rSsHDAC-3, respectively.

Techniques: