Journal: BMC Developmental Biology

Article Title: Histone deacetylase expression patterns in developing murine optic nerve

doi: 10.1186/1471-213X-14-30

Figure Lengend Snippet: Concentration of antibodies used for IHC and Westerns

Article Snippet: Mouse Anti-HDAC4 (Cat# NBP2–22151) , Novus Biologicals (Littleton, Co) , 1:100 , 1:1500 , Abcam (Cambridge, MA).

Techniques: Concentration Assay

Journal: Environmental Health Perspectives

Article Title: E2F1-Mediated FOS Induction in Arsenic Trioxide–Induced Cellular Transformation: Effects of Global H3K9 Hypoacetylation and Promoter-Specific Hyperacetylation in Vitro

doi: 10.1289/ehp.1408302

Figure Lengend Snippet: Histone acetylation analysis of HEK293T and UROtsa cells, identification of H3K9 acetylation control, and mRNA expression analysis of HDAC2 and PCAF in cells treated with As 2 O 3 for 3 or 72 hr. ( A , B ) Histone acetylation (mean ± SD) of H3K9, H4K12, and H4K16 in HEK293T ( A ) and UROtsa ( B ) cells, analyzed by immunoblotting and densitometric analysis (see “Materials and Methods” for details). ( C ) H3K9 acetylation status analyzed by immunoblotting after transfection of HEK293T cells with a no-insert control (NOI), HDAC1, HDAC2, HDAC3, or HDAC4, followed by histone extraction to examine the relative enzymatic effect of these HDACs on this residue; blots were quantified by densitometric analysis and values shown represent three independent experiments. ( D ) Representative immunoblot showing H3K9 acetylation (H3K9ac) status after transfection of HEK293T cells with a NOI or PCAF followed by a 3-hr exposure to 0.5 μM or 2.5 μM As 2 O 3 . ( E , F ) Relative mRNA expression ratios of HDAC2 ( E ) and PCAF ( F ), expressed as fold change for 3 and 72 hr. Values presented are mean ± SD. * p < 0.05 compared with the untreated control [ n = 3 ( B,E ); n = 4 ( F at 3 hr)] calculated by two-tailed Student’s t ‑test.

Article Snippet: We used the following plasmids for transfections: HDAC1 (plasmid 13820), HDAC3 (plasmid 13819), and HDAC4 (plasmid 13821; all from Addgene).

Techniques: Control, Expressing, Western Blot, Transfection, Extraction, Residue, Two Tailed Test

Journal: Muscle & Nerve

Article Title: Leucine Supplementation Counteracts the Atrophic Effects of HDAC4 in Rat Skeletal Muscle Submitted to Hindlimb Immobilization

doi: 10.1002/mus.28411

Figure Lengend Snippet: Soleus muscle morphometric analysis after 7 days of hind limb immobilization and leucine supplementation in animals that overexpressed HDAC4. (A) Representative immunofluorescence photomicrographs of soleus muscle, laminin (green) and DAPI (blue, used for nuclei identification) (scale bar 100 μm) and hematoxylin–eosin staining (scale bar 50 μm). The asterisk corresponds to the same fiber in both HE and immunofluorescence technique. (B) Soleus muscle mass, (C) whole muscle CSA, (D) fiber CSA and (E) fiber distribution from EV, pCMVHDAC4, Imm/pCMVHDAC4 and Imm + Leu/pCMVHDAC4 groups. Data are expressed as mean ± SEM. Statistical analysis included the two‐way ANOVA test followed by Tukey's post hoc . * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 ( n = 5 per group).

Article Snippet: After 30 min, muscles were injected with 50 μL of the HDAC4 expression vector (1.0 μg/μL, pCMVHDAC4, Origene CAT#: RR217458, Rockville, MD, USA) or the empty vector (EV) and an electric pulse generator delivered a burst of pulses (6 pulses of 20 ms with 980 ms pause, 25 V).

Techniques: Immunofluorescence, Staining

Journal: Muscle & Nerve

Article Title: Leucine Supplementation Counteracts the Atrophic Effects of HDAC4 in Rat Skeletal Muscle Submitted to Hindlimb Immobilization

doi: 10.1002/mus.28411

Figure Lengend Snippet: HDAC4 protein expression analysis after 7 days of hind limb immobilization and leucine supplementation in animals that overexpressed HDAC4. (A) HDAC4 and GAPDH western blot showing representative bands. (B) Densitometry analysis by using GAPDH protein level as loading control. Data are expressed as mean ± SEM. Statistical analysis included the two‐way ANOVA test followed by Tukey's post hoc . ** p < 0.01 ( n = 5 per group).

Article Snippet: After 30 min, muscles were injected with 50 μL of the HDAC4 expression vector (1.0 μg/μL, pCMVHDAC4, Origene CAT#: RR217458, Rockville, MD, USA) or the empty vector (EV) and an electric pulse generator delivered a burst of pulses (6 pulses of 20 ms with 980 ms pause, 25 V).

Techniques: Expressing, Western Blot, Control

Journal: Muscle & Nerve

Article Title: Leucine Supplementation Counteracts the Atrophic Effects of HDAC4 in Rat Skeletal Muscle Submitted to Hindlimb Immobilization

doi: 10.1002/mus.28411

Figure Lengend Snippet: Effect of leucine on HDAC4 canonical pathway in animals that overexpressed HDAC4. (A) HDAC4 and (B) Atrogin‐1 mRNA levels were determined by qPCR. (C) Schematic illustrating HDAC4 action in the skeletal muscle fiber nucleus. Cyclophilin A expression was used as housekeeping. Data were expressed as mean ± SEM. Statistical analysis included one‐way ANOVA followed by Tukey's post hoc test. * p < 0.05; ** p < 0.01 ( n = 5 per group).

Article Snippet: After 30 min, muscles were injected with 50 μL of the HDAC4 expression vector (1.0 μg/μL, pCMVHDAC4, Origene CAT#: RR217458, Rockville, MD, USA) or the empty vector (EV) and an electric pulse generator delivered a burst of pulses (6 pulses of 20 ms with 980 ms pause, 25 V).

Techniques: Expressing

Journal: Muscle & Nerve

Article Title: Leucine Supplementation Counteracts the Atrophic Effects of HDAC4 in Rat Skeletal Muscle Submitted to Hindlimb Immobilization

doi: 10.1002/mus.28411

Figure Lengend Snippet: HDAC4 nuclei localization after 7 days of hind limb immobilization and leucine supplementation in animals that overexpressed HDAC4. (A) Representative immunofluorescence photomicrographs of HDAC4 in soleus muscle, HDAC4 (red), DAPI (blue, used for nuclei identification) (scale bar 100 μm). (B) Number of positive HDAC4 nuclei per total nuclei (%). Data were expressed as mean ± SEM. Statistical analysis included two‐way ANOVA followed by Tukey's post hoc test. * p < 0.05; ** p < 0.01 ( n = 5 per group).

Article Snippet: After 30 min, muscles were injected with 50 μL of the HDAC4 expression vector (1.0 μg/μL, pCMVHDAC4, Origene CAT#: RR217458, Rockville, MD, USA) or the empty vector (EV) and an electric pulse generator delivered a burst of pulses (6 pulses of 20 ms with 980 ms pause, 25 V).

Techniques: Immunofluorescence

Journal: International journal of biological sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis.

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: Figure 2. HDAC4 is responsible for deacetylation of GAC at K311. A, B Indicated plasmids were transfected into H1299 cells. Interaction between HDAC4 and GAC was detected by immunoprecipitation and western blot. WCL: whole cell lysate. C Indicated plasmids were transfected into H1299 cells. The protein expression was determined by western blot and glutaminase activity assay was performed. D Indicated siRNAs were transfected into H1299 cells. The protein expression was determined by western blot and glutaminase activity assay was performed. E Indicated plasmids and siRNAs were transfected into H1299 cells and glutaminase activity assay was performed. F The mitochondrial and cytosolic proteins in H1299 cells were separated and the location of HDAC4 and GAC was determined by western blot. VDAC was used as a marker of mitochondrial proteins and GAPDH was used as a marker of cytosolic proteins. Data are showed as mean ± SD, n=3. **P < 0.01, ns P >0.05.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques: Transfection, Immunoprecipitation, Western Blot, Expressing, Activity Assay, Marker

Journal: International journal of biological sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis.

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: Figure 3. HDAC4 promoted cell proliferation and migration in NSCLC cells. A, B Indicated plasmids were transfected into H1299 (A) and A549 (B) cells and cell growth assay was performed. C, D Indicated siRNAs were transfected into H1299 (C) and A549 (D) cells and cell growth assay was performed. E, F Indicated siRNAs and plasmids were transfected into H1299 (E) and A549 (F) cells and cell growth assay was performed. Western blot assay was performed to confirm the transfection efficiency. G, H Indicated siRNAs were transfected into H1299 cells (G) and H292 cells (H) and cell wound healing assay was performed (scale bar: 500 µm, magnification: 100×). I F-actin staining assay. Indicated siRNAs were transfected into H1299 cells. 48 h later, cells were stained with phalloidin and DAPI. Scale bar=20 µm. Data are showed as mean ± SD, n=3. **P<0.01, ***P<0.001.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques: Migration, Transfection, Growth Assay, Western Blot, Wound Healing Assay, Staining

Journal: International journal of biological sciences

Article Title: Deacetylation of Glutaminase by HDAC4 contributes to Lung Cancer Tumorigenesis.

doi: 10.7150/ijbs.69882

Figure Lengend Snippet: Figure 8. A working model of GAC acetylation in NSCLC. A working model depicting the molecular mechanism of HDAC4 mediated GAC deacetylation and TRIM21 mediated GAC ubiquitination to regulation tumorigenesis in NSCLC.

Article Snippet: The HDAC4 siRNAs (OriGene, SR306523) and TRIM21 siRNAs (OriGene, SR304594) were purchased from OriGene.

Techniques: Ubiquitin Proteomics