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anti hdac 2  (Cell Signaling Technology Inc)


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    Structured Review

    Cell Signaling Technology Inc anti hdac 2
    Anti Hdac 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 136 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+hdac+2/HDAC2+Rabbit+mAb/pm41369384-91-14-28
    Average 95 stars, based on 136 article reviews
    anti hdac 2 - by Bioz Stars, 2026-11
    95/100 stars

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    Related Articles

    Polyacrylamide Gel Electrophoresis:

    Article Title: Changes in Class I and IIb HDACs by δ-Opioid in Chronic Rat Glaucoma Model
    Article Snippet: .. Equivalent amounts of protein (10–20 μg) from retina lysates were separated in a Nu PAGE 10% Bis-Tris gel (Thermo Fisher Scientific, Carlsbad, CA, USA) and proteins were transferred to nitrocellulose membranes, as described earlier., The membranes were blocked with 5% non-fat dry milk for 1 hour followed by incubation with primary antibodies (anti-HDAC 1 [Cell Signaling Technology; Cat # D5C6U], anti-HDAC 2 [Cell Signaling Technology; Cat # D6S5P], anti-HDAC 3 [Cell Signaling Technology; Cat # 7G6C5], or anti-HDAC 6 [Cell Signaling Technology; Cat # D21B10] at 1:1000 dilutions, or β-actin [Sigma Aldrich; Cat # A5316] at 1:4000 dilutions) for 16 hours at 4°C. .. After washing, the membranes were incubated for 1 hour at room temperature with the appropriate secondary antibodies (Anti-rabbit IgG, HRP-linked; Cell Signaling Technology; Cat # 7074; anti-mouse IgG, HRP-linked; Cell Signaling Technology; Cat # 7076 dilution 1:5000).

    Incubation:

    Article Title: Changes in Class I and IIb HDACs by δ-Opioid in Chronic Rat Glaucoma Model
    Article Snippet: .. Equivalent amounts of protein (10–20 μg) from retina lysates were separated in a Nu PAGE 10% Bis-Tris gel (Thermo Fisher Scientific, Carlsbad, CA, USA) and proteins were transferred to nitrocellulose membranes, as described earlier., The membranes were blocked with 5% non-fat dry milk for 1 hour followed by incubation with primary antibodies (anti-HDAC 1 [Cell Signaling Technology; Cat # D5C6U], anti-HDAC 2 [Cell Signaling Technology; Cat # D6S5P], anti-HDAC 3 [Cell Signaling Technology; Cat # 7G6C5], or anti-HDAC 6 [Cell Signaling Technology; Cat # D21B10] at 1:1000 dilutions, or β-actin [Sigma Aldrich; Cat # A5316] at 1:4000 dilutions) for 16 hours at 4°C. .. After washing, the membranes were incubated for 1 hour at room temperature with the appropriate secondary antibodies (Anti-rabbit IgG, HRP-linked; Cell Signaling Technology; Cat # 7074; anti-mouse IgG, HRP-linked; Cell Signaling Technology; Cat # 7076 dilution 1:5000).

    Article Title: Altérations fonctionnelles des cellules dentritiques suivant une brûlure sévère
    Article Snippet: .. Membranes were blocked and incubated overnight at 4°C with the following primary antibodies: anti-HDAC—l, anti-HDAC-2 and anti-HDAC 3 (rabbit polyclonal; Cell Signaling, Pickering, ONT, Canada) and phospho-p38 MAPK (rabbit polyclonal; BioSource, Camarillo, CA, USA). .. The membranes were subsequently incubated with a second antibody HRP- conjugated anti-rabbit lgG (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and visualized using Western Chemiluminescence ECL Plus detection reagents (Amersham Biosciences, Baie d’Urfé, QC, Canada) according to the manufacturer’s protocol.



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    Immunohistochemical expression of <t>HDAC-2</t> ( A , D ), HDAC-4 ( B , E ), and HDAC-5 ( C , F ) in epithelial ovarian carcinomas. ( A ) A high-grade serous ovarian carcinoma with nuclear HDAC-2 expression (×200). ( B ) A high-grade serous ovarian carcinoma with cytoplasmic HDAC-4 expression (×200). ( C ) A high-grade serous ovarian carcinoma with cytoplasmic HDAC-5 expression (×400). ( D ) A high-grade serous ovarian carcinoma with nuclear HDAC-2 expression (×400). ( E ) A high-grade serous ovarian carcinoma with cytoplasmic HDAC-4 expression (×400). ( F ) A high-grade serous ovarian carcinoma with nuclear and cytoplasmic HDAC-4 expression (×400).
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    Image Search Results


    HDAC2 enhances antimicrobial activities against E.coli in mice. (A) Bacterial loads in the blood were calculated in HDAC2-knockout and wild type mice. (A)The picture of the bacterial clones was taken by BIO-RAD ChemiDoc“MP Imaging System. (B)The number of colony-forming units (CFU) in the blood of mice was calculated. HDAC2 WT and HDAC2 KO mice were administrated with E. coli- GFP (5 × 10 7 ) by intravenous Injection. Blood (20 μl) was taken from the tail vein of the mice, applied to the pretreated solid MHA medium, and incubated at 37 °C for 20 h. Count the colonies on the culture medium and take pictures. The experiments were performed in quintuplicate, data are presented as the means ± SEM. of independent experiments. *P < 0.05, **P < 0.01 (two-tailed Student’s t -test). (B) Bacterial loads in the kidneys were calculated in HDAC2-knockout and wild type mice. The picture of kidneys from HDAC2 WT and HDAC2 KO mice were taken. The number of colony-forming units (CFU) in the kidneys of mice was calculated. The kidneys from HDAC2 KO and HDAC2 WT mice were weighed, ground, spread on the MHA plates, incubated for 16–24 h at 37 °C, and then bacterial clones were counted. The experiments were performed in quintuplicate, data are presented as the means ± SEM. of independent experiments. *P < 0.05, **P < 0.01 (two-tailed Student’s t -test). n = 5 mice per group. (C) H&E staining of livers and kidneys. The arrows indicate damage section of tissue. (D) Serum CRP and PCT levels of mice with E. coli- GFP bacteremia at 24 h. The CRP and PCT were measured by ELISA. Data are shown as mean ± SEM. *p < 0.05; n = 5 mice per group. ns, not significant.

    Journal: Journal of Advanced Research

    Article Title: HDAC2 enhances the antimicrobial activity of neutrophils by promoting the formation of neutrophil extracellular traps (NETs) in sepsis

    doi: 10.1016/j.jare.2025.08.041

    Figure Lengend Snippet: HDAC2 enhances antimicrobial activities against E.coli in mice. (A) Bacterial loads in the blood were calculated in HDAC2-knockout and wild type mice. (A)The picture of the bacterial clones was taken by BIO-RAD ChemiDoc“MP Imaging System. (B)The number of colony-forming units (CFU) in the blood of mice was calculated. HDAC2 WT and HDAC2 KO mice were administrated with E. coli- GFP (5 × 10 7 ) by intravenous Injection. Blood (20 μl) was taken from the tail vein of the mice, applied to the pretreated solid MHA medium, and incubated at 37 °C for 20 h. Count the colonies on the culture medium and take pictures. The experiments were performed in quintuplicate, data are presented as the means ± SEM. of independent experiments. *P < 0.05, **P < 0.01 (two-tailed Student’s t -test). (B) Bacterial loads in the kidneys were calculated in HDAC2-knockout and wild type mice. The picture of kidneys from HDAC2 WT and HDAC2 KO mice were taken. The number of colony-forming units (CFU) in the kidneys of mice was calculated. The kidneys from HDAC2 KO and HDAC2 WT mice were weighed, ground, spread on the MHA plates, incubated for 16–24 h at 37 °C, and then bacterial clones were counted. The experiments were performed in quintuplicate, data are presented as the means ± SEM. of independent experiments. *P < 0.05, **P < 0.01 (two-tailed Student’s t -test). n = 5 mice per group. (C) H&E staining of livers and kidneys. The arrows indicate damage section of tissue. (D) Serum CRP and PCT levels of mice with E. coli- GFP bacteremia at 24 h. The CRP and PCT were measured by ELISA. Data are shown as mean ± SEM. *p < 0.05; n = 5 mice per group. ns, not significant.

    Article Snippet: Concentrations of interleukin-1 beta (IL-1β), interleukin-6 (IL-6), histone deacetylase 2 (HDAC2), PCT, CRP and myeloperoxidase (MPO) in mouse serum were quantified using their respective enzyme-linked immunosorbent assay (ELISA) kits, following the protocols provided by the manufacturer (Boster Biological Technology, Wuhan, China).

    Techniques: Knock-Out, Clone Assay, Imaging, Injection, Incubation, Two Tailed Test, Staining, Enzyme-linked Immunosorbent Assay

    Figure 1. Sp7-Cre; Hdac4fl/fltransgenic mice were successfully generated and developed dwarfism with skeletal dysplasia. A, schematic representation and breeding strategies of the Sp7-Cre; Hdac4fl/fltransgenic mice and Acan-CreERT2; Hdac4fl/fltransgenic mice. B, the Gross phenotype and representative X-ray images of 2-month-old male control and Sp7-Cre; Hdac4fl/fl. Mutants exhibit a dwarf body and short limbs. Purple or blue boxes indicate magnified areas, showing short hindlimbs, and osteopenia (arrow). Black bars represent 2 cm, white bars represent 2 cm. C, measurement of body weight and length of 2-month-old mice (n = 4). Measurement of femur length (% of normal femur length) (n = 3), the length of tibia (% of normal tibia length) (n = 3). Between-group comparisons utilized Student’s t-tests. The p-values for statistical comparisons between groups are shown in the Figure. D, immunohistochemistry staining of Hdac4 on paraffin sections of 2-month-old Hdac4fl/flor Sp7-Cre; Hdac4fl/fltibias. The results showed that Hdac4 protein was weakly expressed both in the tibial metaphysis, subchondral trabeculae, and tibial cortex. Bar represents 50 mm or 20 mm. E, Western blots and RT-qPCR show the protein and mRNA expression levels of Hdac4 and ALP in tibias derived from 2-month-old Hdac4fl/flor Sp7-Cre; Hdac4fl/fl, respectively. b-actin served as an internal control. Data are the mean ± SD, n = 3. Table S1 presents the primer sequences used for the RT-qPCR.

    Journal: The Journal of biological chemistry

    Article Title: Loss of Hdac4 in osteoprogenitors impairs postnatal trabecular and cortical bone formation, resulting in a dwarfism and osteopenia phenotype in mice.

    doi: 10.1016/j.jbc.2024.107941

    Figure Lengend Snippet: Figure 1. Sp7-Cre; Hdac4fl/fltransgenic mice were successfully generated and developed dwarfism with skeletal dysplasia. A, schematic representation and breeding strategies of the Sp7-Cre; Hdac4fl/fltransgenic mice and Acan-CreERT2; Hdac4fl/fltransgenic mice. B, the Gross phenotype and representative X-ray images of 2-month-old male control and Sp7-Cre; Hdac4fl/fl. Mutants exhibit a dwarf body and short limbs. Purple or blue boxes indicate magnified areas, showing short hindlimbs, and osteopenia (arrow). Black bars represent 2 cm, white bars represent 2 cm. C, measurement of body weight and length of 2-month-old mice (n = 4). Measurement of femur length (% of normal femur length) (n = 3), the length of tibia (% of normal tibia length) (n = 3). Between-group comparisons utilized Student’s t-tests. The p-values for statistical comparisons between groups are shown in the Figure. D, immunohistochemistry staining of Hdac4 on paraffin sections of 2-month-old Hdac4fl/flor Sp7-Cre; Hdac4fl/fltibias. The results showed that Hdac4 protein was weakly expressed both in the tibial metaphysis, subchondral trabeculae, and tibial cortex. Bar represents 50 mm or 20 mm. E, Western blots and RT-qPCR show the protein and mRNA expression levels of Hdac4 and ALP in tibias derived from 2-month-old Hdac4fl/flor Sp7-Cre; Hdac4fl/fl, respectively. b-actin served as an internal control. Data are the mean ± SD, n = 3. Table S1 presents the primer sequences used for the RT-qPCR.

    Article Snippet: Subsequent to incubation with primary antibodies against Hdac4 (Abcolnal, A0239 or Cell Signaling Technology, 7628), Col1a1, ALP, osteocalcin, bactin, and the corresponding HRP-labeled secondary antibody, protein bands were detected using a highly sensitive ECL chemiluminescence kit (Boster) and visualized with a Bio-Rad imaging system.

    Techniques: Generated, Control, Immunohistochemistry, Staining, Western Blot, Quantitative RT-PCR, Expressing, Derivative Assay

    Figure 2. Loss of Hdac4 in Sp7-expressing osteoprogenitors resulted in reduced cortical and cancellous bone formation in the 2 months tibias. A, representative images of Masson’s trichrome staining showed that sparse trabecular bone and thin cortical bone were observed in Sp7-Cre; Hdac4fl/flmice at 2 months. Scale bars are shown in the correspondence diagram. Black arrows or arrowheads indicate trabeculae and cortex of the long bone in two groups of mice, respectively. B, representative X-ray images showing whole tibias (upper panel); representative micro-CT images of tibias from 2-month-old mice (lower panel) show the proximal tibia (top; scale bars represent 1 mm), trabecular bone of the tibia metaphysis (middle), and cortical (bottom). C, indicators for evaluating the spatial morphologic structure of bone trabeculae (i–iv): bone volume as a percentage of total volume (BV/TV, %), trabecular thickness (Tb. Th, mm), number of trabeculae (Tb. N, 1/mm), and trabecular separation (Tb. Sp, 1/mm). Cortical bone was analyzed as follows (v–viii): total cortical bone area (Tt. Ar, mm2), cortical bone area (Ct. Ar, mm2), cortical bone thickness (Ct. Th, mm), and cortical bone area as a percentage of total area (Ct. Ar/Tt. Ar, %). Hdac4fl/fl

    Journal: The Journal of biological chemistry

    Article Title: Loss of Hdac4 in osteoprogenitors impairs postnatal trabecular and cortical bone formation, resulting in a dwarfism and osteopenia phenotype in mice.

    doi: 10.1016/j.jbc.2024.107941

    Figure Lengend Snippet: Figure 2. Loss of Hdac4 in Sp7-expressing osteoprogenitors resulted in reduced cortical and cancellous bone formation in the 2 months tibias. A, representative images of Masson’s trichrome staining showed that sparse trabecular bone and thin cortical bone were observed in Sp7-Cre; Hdac4fl/flmice at 2 months. Scale bars are shown in the correspondence diagram. Black arrows or arrowheads indicate trabeculae and cortex of the long bone in two groups of mice, respectively. B, representative X-ray images showing whole tibias (upper panel); representative micro-CT images of tibias from 2-month-old mice (lower panel) show the proximal tibia (top; scale bars represent 1 mm), trabecular bone of the tibia metaphysis (middle), and cortical (bottom). C, indicators for evaluating the spatial morphologic structure of bone trabeculae (i–iv): bone volume as a percentage of total volume (BV/TV, %), trabecular thickness (Tb. Th, mm), number of trabeculae (Tb. N, 1/mm), and trabecular separation (Tb. Sp, 1/mm). Cortical bone was analyzed as follows (v–viii): total cortical bone area (Tt. Ar, mm2), cortical bone area (Ct. Ar, mm2), cortical bone thickness (Ct. Th, mm), and cortical bone area as a percentage of total area (Ct. Ar/Tt. Ar, %). Hdac4fl/fl

    Article Snippet: Subsequent to incubation with primary antibodies against Hdac4 (Abcolnal, A0239 or Cell Signaling Technology, 7628), Col1a1, ALP, osteocalcin, bactin, and the corresponding HRP-labeled secondary antibody, protein bands were detected using a highly sensitive ECL chemiluminescence kit (Boster) and visualized with a Bio-Rad imaging system.

    Techniques: Expressing, Staining, Micro-CT

    Figure 3. Loss of Hdac4 in Sp7-expressing osteoprogenitors mildly affects postnatal bone development in pre-weaned mice but allows the formation of growth plates and secondary ossification centers. A, serial analyses of postnatal tibia phenotype using whole skeleton staining and X-ray image in Hdac4fl/fland Sp7-Cre; Hdac4fl/flmice at P1, P7, and P14. Bars represent 2 mm. B, measurement of body length in the mice of different ages. n = 4. Data are the mean ± SD, the p- values for statistical comparisons between groups are shown in the Figure. C, H&E stain and Von Kossa stain of longitudinal tibia sections from Hdac4fl/fl, Sp7-Cre; Hdac4fl/fland Acan-CreERT2; Hdac4fl/flmice at P7, P14. As shown, Sp7-Cre; Hdac4fl/flmice displayed delayed onset of cartilaginous canal at P7, weak mineral deposition in primary ossification centers of tibias at P14, but thinner growth plates and strong mineralization of primary sponges were observed in the corre- sponding region of the Acan-CreERT2; Hdac4fl/fl. D, H&E stain showed that a decrease in epiphyseal osteogenesis were observed in Sp7-Cre; Hdac4fl/flmice, but a complete loss of the growth plate, accelerated ossification of the epiphysis in Acan-CreERT2; Hdac4fl/fltibias at P21. Bars represent 200 mm in the upper panels; bar represents 50 mm in the magnified panels.

    Journal: The Journal of biological chemistry

    Article Title: Loss of Hdac4 in osteoprogenitors impairs postnatal trabecular and cortical bone formation, resulting in a dwarfism and osteopenia phenotype in mice.

    doi: 10.1016/j.jbc.2024.107941

    Figure Lengend Snippet: Figure 3. Loss of Hdac4 in Sp7-expressing osteoprogenitors mildly affects postnatal bone development in pre-weaned mice but allows the formation of growth plates and secondary ossification centers. A, serial analyses of postnatal tibia phenotype using whole skeleton staining and X-ray image in Hdac4fl/fland Sp7-Cre; Hdac4fl/flmice at P1, P7, and P14. Bars represent 2 mm. B, measurement of body length in the mice of different ages. n = 4. Data are the mean ± SD, the p- values for statistical comparisons between groups are shown in the Figure. C, H&E stain and Von Kossa stain of longitudinal tibia sections from Hdac4fl/fl, Sp7-Cre; Hdac4fl/fland Acan-CreERT2; Hdac4fl/flmice at P7, P14. As shown, Sp7-Cre; Hdac4fl/flmice displayed delayed onset of cartilaginous canal at P7, weak mineral deposition in primary ossification centers of tibias at P14, but thinner growth plates and strong mineralization of primary sponges were observed in the corre- sponding region of the Acan-CreERT2; Hdac4fl/fl. D, H&E stain showed that a decrease in epiphyseal osteogenesis were observed in Sp7-Cre; Hdac4fl/flmice, but a complete loss of the growth plate, accelerated ossification of the epiphysis in Acan-CreERT2; Hdac4fl/fltibias at P21. Bars represent 200 mm in the upper panels; bar represents 50 mm in the magnified panels.

    Article Snippet: Subsequent to incubation with primary antibodies against Hdac4 (Abcolnal, A0239 or Cell Signaling Technology, 7628), Col1a1, ALP, osteocalcin, bactin, and the corresponding HRP-labeled secondary antibody, protein bands were detected using a highly sensitive ECL chemiluminescence kit (Boster) and visualized with a Bio-Rad imaging system.

    Techniques: Expressing, Staining

    Figure 4. Loss of Hdac4 in Sp7-expressing osteoprogenitors leads to reduced trabecular bone formation and thinner growth plates. A, H&E stain, Safranin O/fast green stain of tibia sections. B, measurement of the thickness of metaphyseal zone and growth plate thickness of 2-month-old tibias. Results are expressed as mean ± SD, the p-values for statistical comparisons between groups are shown in the Figure, n = 5. C, immunohistochemistry analysis of tibia sections using the primary antibody of Col1a1 and Ihh (Indian hedgehog). The boxed regions in the previous row of pictures were magnified in the right pictures, respectively. Scale bar represents 200 mm, 50 mm, 20 mm, respectively. D, frozen sections showed that Cre-GFP was detected as green fluorescence in the tibia of Sp7-Cre; Rosa26-ZsGreen1 and Sp7-Cre; Hdac4fl/fl; Rosa26-ZsGreen1 mice at 2 months. E, tartrate-resistant acid phosphatase (TRAP) staining was performed on the tibial tissue of 2-month-old Hdac4fl/fland Sp7-Cre; Hdac4fl/flmice. F, quantification of the numbers of GFP-positive osteoblasts, TRAP-positive osteoclasts, and percentage of osteoclast surface in the proximal metaphysis. Results are expressed as mean ± SD, n = 4. The p-values for statistical comparisons between groups are shown in corresponding images.

    Journal: The Journal of biological chemistry

    Article Title: Loss of Hdac4 in osteoprogenitors impairs postnatal trabecular and cortical bone formation, resulting in a dwarfism and osteopenia phenotype in mice.

    doi: 10.1016/j.jbc.2024.107941

    Figure Lengend Snippet: Figure 4. Loss of Hdac4 in Sp7-expressing osteoprogenitors leads to reduced trabecular bone formation and thinner growth plates. A, H&E stain, Safranin O/fast green stain of tibia sections. B, measurement of the thickness of metaphyseal zone and growth plate thickness of 2-month-old tibias. Results are expressed as mean ± SD, the p-values for statistical comparisons between groups are shown in the Figure, n = 5. C, immunohistochemistry analysis of tibia sections using the primary antibody of Col1a1 and Ihh (Indian hedgehog). The boxed regions in the previous row of pictures were magnified in the right pictures, respectively. Scale bar represents 200 mm, 50 mm, 20 mm, respectively. D, frozen sections showed that Cre-GFP was detected as green fluorescence in the tibia of Sp7-Cre; Rosa26-ZsGreen1 and Sp7-Cre; Hdac4fl/fl; Rosa26-ZsGreen1 mice at 2 months. E, tartrate-resistant acid phosphatase (TRAP) staining was performed on the tibial tissue of 2-month-old Hdac4fl/fland Sp7-Cre; Hdac4fl/flmice. F, quantification of the numbers of GFP-positive osteoblasts, TRAP-positive osteoclasts, and percentage of osteoclast surface in the proximal metaphysis. Results are expressed as mean ± SD, n = 4. The p-values for statistical comparisons between groups are shown in corresponding images.

    Article Snippet: Subsequent to incubation with primary antibodies against Hdac4 (Abcolnal, A0239 or Cell Signaling Technology, 7628), Col1a1, ALP, osteocalcin, bactin, and the corresponding HRP-labeled secondary antibody, protein bands were detected using a highly sensitive ECL chemiluminescence kit (Boster) and visualized with a Bio-Rad imaging system.

    Techniques: Expressing, Staining, Immunohistochemistry

    Figure 5. Loss of Hdac4 in Sp7-expressing osteoprogenitors leads to reduced expression of major bone matrix proteins and decreased angio- genesis in the metaphyseal region. A–C, the protein expression profiles and gene expression levels of osteoblast and osteoclast markers ALP, osteocalcin, OPG, and SOST were examined using immunohistochemistry and RT-qPCR. Examination of protein expression profiles and gene expression levels of the markers of terminal chondrocyte differentiation (Col10a1, MMP13) using immunohistochemistry (IHC) and RT-qPCR. D, confocal images of immunofluo- rescence staining for Endomucin (Emcn) and Cre-GFP on frozen sections of the proximal tibia at P21; CD31 immunohistochemical staining at P21. E, CD31and VEGFa immunohistochemical staining of the proximal tibia in the experimental and control group at 2 months. F, the mRNA expression of angiogenesis-related genes (CD31,VEGFa) at 2 months was detected using RT-qPCR. n = 3, the p-values are shown in corresponding images. The values of Hdac4fl/flmice were defined as 1. Results are expressed as mean ± SD. Scale bars are 100 mm and 40 mm, respectively.

    Journal: The Journal of biological chemistry

    Article Title: Loss of Hdac4 in osteoprogenitors impairs postnatal trabecular and cortical bone formation, resulting in a dwarfism and osteopenia phenotype in mice.

    doi: 10.1016/j.jbc.2024.107941

    Figure Lengend Snippet: Figure 5. Loss of Hdac4 in Sp7-expressing osteoprogenitors leads to reduced expression of major bone matrix proteins and decreased angio- genesis in the metaphyseal region. A–C, the protein expression profiles and gene expression levels of osteoblast and osteoclast markers ALP, osteocalcin, OPG, and SOST were examined using immunohistochemistry and RT-qPCR. Examination of protein expression profiles and gene expression levels of the markers of terminal chondrocyte differentiation (Col10a1, MMP13) using immunohistochemistry (IHC) and RT-qPCR. D, confocal images of immunofluo- rescence staining for Endomucin (Emcn) and Cre-GFP on frozen sections of the proximal tibia at P21; CD31 immunohistochemical staining at P21. E, CD31and VEGFa immunohistochemical staining of the proximal tibia in the experimental and control group at 2 months. F, the mRNA expression of angiogenesis-related genes (CD31,VEGFa) at 2 months was detected using RT-qPCR. n = 3, the p-values are shown in corresponding images. The values of Hdac4fl/flmice were defined as 1. Results are expressed as mean ± SD. Scale bars are 100 mm and 40 mm, respectively.

    Article Snippet: Subsequent to incubation with primary antibodies against Hdac4 (Abcolnal, A0239 or Cell Signaling Technology, 7628), Col1a1, ALP, osteocalcin, bactin, and the corresponding HRP-labeled secondary antibody, protein bands were detected using a highly sensitive ECL chemiluminescence kit (Boster) and visualized with a Bio-Rad imaging system.

    Techniques: Expressing, Gene Expression, Immunohistochemistry, Quantitative RT-PCR, Staining, Immunohistochemical staining, Control

    Figure 6. Loss of Hdac4 in Sp7-expressing osteoprogenitors inhibited the differentiation and mineralization of osteoblasts in vitro. A, primary osteoblasts were isolated from the calvaria of postnatal days 6 to 7 Hdac4fl/flmice and Sp7-Cre; Hdac4fl/flmice and then cultured for proliferation and differentiation assays. B, relative to the Hdac4fl/flgroup, the protein level of Hdac4 decreased in the primary osteoblasts. C, primary osteoblast from calvarias of Sp7-Cre; Hdac4fl/flmice and control littermates were cultured for 36 h, 72 h in growth medium and then subjected to CCK8 assay. Results are expressed as mean ± SD, the p-values for are shown in corresponding images. D, alizarin red staining of primary osteoblast was performed after 6 and 12 days of culture in osteogenic medium, respectively. E, ALP staining and von Kossa staining of primary osteoblast were performed after 14 days of culture in osteogenic medium, respectively. Scale bars represent 5 mm. F and G, cells were harvested after 8 and 15 days of culture in osteogenic medium and subjected to RT- qPCR assay and Western blot assay to analyse the mRNA and protein levels of Col1a1, Alp, and Osteocalcin, respectively. Results are expressed as mean ± SD, n = 3. The p-values for are shown in corresponding images.

    Journal: The Journal of biological chemistry

    Article Title: Loss of Hdac4 in osteoprogenitors impairs postnatal trabecular and cortical bone formation, resulting in a dwarfism and osteopenia phenotype in mice.

    doi: 10.1016/j.jbc.2024.107941

    Figure Lengend Snippet: Figure 6. Loss of Hdac4 in Sp7-expressing osteoprogenitors inhibited the differentiation and mineralization of osteoblasts in vitro. A, primary osteoblasts were isolated from the calvaria of postnatal days 6 to 7 Hdac4fl/flmice and Sp7-Cre; Hdac4fl/flmice and then cultured for proliferation and differentiation assays. B, relative to the Hdac4fl/flgroup, the protein level of Hdac4 decreased in the primary osteoblasts. C, primary osteoblast from calvarias of Sp7-Cre; Hdac4fl/flmice and control littermates were cultured for 36 h, 72 h in growth medium and then subjected to CCK8 assay. Results are expressed as mean ± SD, the p-values for are shown in corresponding images. D, alizarin red staining of primary osteoblast was performed after 6 and 12 days of culture in osteogenic medium, respectively. E, ALP staining and von Kossa staining of primary osteoblast were performed after 14 days of culture in osteogenic medium, respectively. Scale bars represent 5 mm. F and G, cells were harvested after 8 and 15 days of culture in osteogenic medium and subjected to RT- qPCR assay and Western blot assay to analyse the mRNA and protein levels of Col1a1, Alp, and Osteocalcin, respectively. Results are expressed as mean ± SD, n = 3. The p-values for are shown in corresponding images.

    Article Snippet: Subsequent to incubation with primary antibodies against Hdac4 (Abcolnal, A0239 or Cell Signaling Technology, 7628), Col1a1, ALP, osteocalcin, bactin, and the corresponding HRP-labeled secondary antibody, protein bands were detected using a highly sensitive ECL chemiluminescence kit (Boster) and visualized with a Bio-Rad imaging system.

    Techniques: Expressing, In Vitro, Isolation, Cell Culture, Control, CCK-8 Assay, Staining, Quantitative RT-PCR, Western Blot

    Figure 7. Loss of Hdac4 in Sp7-expressing osteoprogenitors alters the gene expression profile and affects multiple biological processes. A, the bar chart shows the total differentially expressed genes (DEGs), upregulated DEGs, and downregulated DEGs. B, volcano plot of the differentially expressed genes (DEGs) for Hdac4fl/flversus Sp7-Cre; Hdac4fl/fl. C, gene Ontology (GO) functional enrichment analysis of all the DEGs for Hdac4fl/fland Sp7-Cre; Hdac4fl/flmice. D, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of all the differentially expressed genes (DEGs) for Hdac4fl/fland Sp7-Cre; Hdac4fl/flmice. E, immunohistochemistry analysis of tibia sections from Hdac4fl/fland Sp7-Cre; Hdac4fl/flat 2-month-old using the primary antibody of SOX9 and Col2a1. F, RNA isolated from control and mutant tibias at 2-month-old was subjected to RT-qPCR assay, and gene expression analysis of SOX9 and Col2a1 was conducted. All target genes’ mRNA levels were normalized to Gapdh mRNA. Results are expressed as mean ± SD, n = 3. The p-values for are shown in corresponding images.

    Journal: The Journal of biological chemistry

    Article Title: Loss of Hdac4 in osteoprogenitors impairs postnatal trabecular and cortical bone formation, resulting in a dwarfism and osteopenia phenotype in mice.

    doi: 10.1016/j.jbc.2024.107941

    Figure Lengend Snippet: Figure 7. Loss of Hdac4 in Sp7-expressing osteoprogenitors alters the gene expression profile and affects multiple biological processes. A, the bar chart shows the total differentially expressed genes (DEGs), upregulated DEGs, and downregulated DEGs. B, volcano plot of the differentially expressed genes (DEGs) for Hdac4fl/flversus Sp7-Cre; Hdac4fl/fl. C, gene Ontology (GO) functional enrichment analysis of all the DEGs for Hdac4fl/fland Sp7-Cre; Hdac4fl/flmice. D, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of all the differentially expressed genes (DEGs) for Hdac4fl/fland Sp7-Cre; Hdac4fl/flmice. E, immunohistochemistry analysis of tibia sections from Hdac4fl/fland Sp7-Cre; Hdac4fl/flat 2-month-old using the primary antibody of SOX9 and Col2a1. F, RNA isolated from control and mutant tibias at 2-month-old was subjected to RT-qPCR assay, and gene expression analysis of SOX9 and Col2a1 was conducted. All target genes’ mRNA levels were normalized to Gapdh mRNA. Results are expressed as mean ± SD, n = 3. The p-values for are shown in corresponding images.

    Article Snippet: Subsequent to incubation with primary antibodies against Hdac4 (Abcolnal, A0239 or Cell Signaling Technology, 7628), Col1a1, ALP, osteocalcin, bactin, and the corresponding HRP-labeled secondary antibody, protein bands were detected using a highly sensitive ECL chemiluminescence kit (Boster) and visualized with a Bio-Rad imaging system.

    Techniques: Expressing, Gene Expression, Functional Assay, Immunohistochemistry, Isolation, Control, Mutagenesis, Quantitative RT-PCR

    Gene expression time-course analysis confirms alterations in the expression of Class II HDAC isoforms during DPC mineralisation. Relative gene expression, using quantitative real-time PCR, of DPCs cultured in a mineralisation medium was analysed relative to day 0 expression and presented as fold change. Day 0 corresponds to a 1-fold change on each panel graph. Significant differences related to HDAC-4 which was significantly increased ( D ) while HDAC-6 was decreased at day 14 ( F ). ( A ) HDAC-1 ; ( B ) HDAC-2 ; ( C ) HDAC-3 ; ( D ) HDAC-4 ; ( E ) HDAC-5 ; and ( F ) HDAC-6 . Β-actin was used as an internal control. Error bars represent means ± SEM of three independent biological samples carried out in triplicate. One-way analysis of variance (ANOVA) and post hoc Tukey’s test were used to ascertain statistical significance at p < 0.05 *.

    Journal: International Journal of Molecular Sciences

    Article Title: Dynamic Alterations in Acetylation and Modulation of Histone Deacetylase Expression Evident in the Dentine–Pulp Complex during Dentinogenesis

    doi: 10.3390/ijms25126569

    Figure Lengend Snippet: Gene expression time-course analysis confirms alterations in the expression of Class II HDAC isoforms during DPC mineralisation. Relative gene expression, using quantitative real-time PCR, of DPCs cultured in a mineralisation medium was analysed relative to day 0 expression and presented as fold change. Day 0 corresponds to a 1-fold change on each panel graph. Significant differences related to HDAC-4 which was significantly increased ( D ) while HDAC-6 was decreased at day 14 ( F ). ( A ) HDAC-1 ; ( B ) HDAC-2 ; ( C ) HDAC-3 ; ( D ) HDAC-4 ; ( E ) HDAC-5 ; and ( F ) HDAC-6 . Β-actin was used as an internal control. Error bars represent means ± SEM of three independent biological samples carried out in triplicate. One-way analysis of variance (ANOVA) and post hoc Tukey’s test were used to ascertain statistical significance at p < 0.05 *.

    Article Snippet: The following antibodies were used: anti-HDAC-1, anti-HDAC-2, anti-HDAC-3, anti-HDAC-4, anti-HDAC-5, anti-HDAC-6 (all 1:100, Abcam, Cambridge, UK), or normal immunoglobulin G (IgG) as negative control; this was performed overnight at 4 °C.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Control

    Time-course analysis of HDAC expression during DPC mineralisation showing altered expression of HDAC isoforms. HDAC expression of DPCs cultured in a mineralisation medium. HDAC-1, -2, and -4 showed increased expression until day 7 and then decreased ( A – C ) while HDAC-6 remained relatively high after day 4 ( D ). Images are representative of Western blotting result from each HDAC isoform (upper) and its internal control (β-actin; lower). Expression levels were normalised and presented as fold changes against day 0 expression. ( A ) HDAC-1; ( B ) HDAC-2; ( C ) HDAC-4; ( D ) HDAC-6. The expression levels of HDAC-3 and -5 were too weak to evaluate with all the samples tested ( E , F ). Error bars represent means ± SEM of at least three independent biological replicates. Alizarin red staining of DPC mineralisation-induced culture shows the mineralisation starts around day 7 and continues until day 21 ( G ).

    Journal: International Journal of Molecular Sciences

    Article Title: Dynamic Alterations in Acetylation and Modulation of Histone Deacetylase Expression Evident in the Dentine–Pulp Complex during Dentinogenesis

    doi: 10.3390/ijms25126569

    Figure Lengend Snippet: Time-course analysis of HDAC expression during DPC mineralisation showing altered expression of HDAC isoforms. HDAC expression of DPCs cultured in a mineralisation medium. HDAC-1, -2, and -4 showed increased expression until day 7 and then decreased ( A – C ) while HDAC-6 remained relatively high after day 4 ( D ). Images are representative of Western blotting result from each HDAC isoform (upper) and its internal control (β-actin; lower). Expression levels were normalised and presented as fold changes against day 0 expression. ( A ) HDAC-1; ( B ) HDAC-2; ( C ) HDAC-4; ( D ) HDAC-6. The expression levels of HDAC-3 and -5 were too weak to evaluate with all the samples tested ( E , F ). Error bars represent means ± SEM of at least three independent biological replicates. Alizarin red staining of DPC mineralisation-induced culture shows the mineralisation starts around day 7 and continues until day 21 ( G ).

    Article Snippet: The following antibodies were used: anti-HDAC-1, anti-HDAC-2, anti-HDAC-3, anti-HDAC-4, anti-HDAC-5, anti-HDAC-6 (all 1:100, Abcam, Cambridge, UK), or normal immunoglobulin G (IgG) as negative control; this was performed overnight at 4 °C.

    Techniques: Expressing, Cell Culture, Western Blot, Control, Staining

    Class I HDAC isoforms exhibit altered expression in postnatal and adult molar teeth. Comparison of Class I HDAC expression in rat molars at postnatal 5 days (first maxillary molar, ( A , C , E )) and 8 weeks (third maxillary molar, ( B , D , F )). ( A , B ) HDAC-1 is highly expressed in postnatal dental pulp, odontoblasts (( A )—black arrows), and ameloblasts compared to negative control (Aiii) but reduced in adult teeth ( B ). ( C , D ) Expression of HDAC-2 was low, not being evident in both developing ( C ) and mature ( D ) teeth. ( E , F ) Meanwhile, HDAC-3 expression was also low in both developing ( E ) and mature ( F ) teeth. HDAC-1 to 3 antibody-positive cells demarcated in brown in individual panels. ( Aii – Fii ) represent higher magnifications of the black boxes in ( Ai – Fi ), respectively. ( Aiv – Fiv ) represent higher magnifications of the black boxes in ( Aii – Fii ), respectively. Inset images ( Aiii – Fiii ) represent negative controls of each respective experiment. Scale bars = ( Ai – Fi ) 200 μm (original mag. ×2.5), ( Aii – Fii , Aiii – Fiii ) 50 µm (original mag. ×10), and ( Aiv – Fiv ) 10 µm (original mag. ×40). Abbreviations: P, pulp tissue; Dn, dentine; Od, odontoblasts; En, enamel. Multiple sections were examined from 5 independent rats at 5 days and 8 weeks, respectively.

    Journal: International Journal of Molecular Sciences

    Article Title: Dynamic Alterations in Acetylation and Modulation of Histone Deacetylase Expression Evident in the Dentine–Pulp Complex during Dentinogenesis

    doi: 10.3390/ijms25126569

    Figure Lengend Snippet: Class I HDAC isoforms exhibit altered expression in postnatal and adult molar teeth. Comparison of Class I HDAC expression in rat molars at postnatal 5 days (first maxillary molar, ( A , C , E )) and 8 weeks (third maxillary molar, ( B , D , F )). ( A , B ) HDAC-1 is highly expressed in postnatal dental pulp, odontoblasts (( A )—black arrows), and ameloblasts compared to negative control (Aiii) but reduced in adult teeth ( B ). ( C , D ) Expression of HDAC-2 was low, not being evident in both developing ( C ) and mature ( D ) teeth. ( E , F ) Meanwhile, HDAC-3 expression was also low in both developing ( E ) and mature ( F ) teeth. HDAC-1 to 3 antibody-positive cells demarcated in brown in individual panels. ( Aii – Fii ) represent higher magnifications of the black boxes in ( Ai – Fi ), respectively. ( Aiv – Fiv ) represent higher magnifications of the black boxes in ( Aii – Fii ), respectively. Inset images ( Aiii – Fiii ) represent negative controls of each respective experiment. Scale bars = ( Ai – Fi ) 200 μm (original mag. ×2.5), ( Aii – Fii , Aiii – Fiii ) 50 µm (original mag. ×10), and ( Aiv – Fiv ) 10 µm (original mag. ×40). Abbreviations: P, pulp tissue; Dn, dentine; Od, odontoblasts; En, enamel. Multiple sections were examined from 5 independent rats at 5 days and 8 weeks, respectively.

    Article Snippet: The following antibodies were used: anti-HDAC-1, anti-HDAC-2, anti-HDAC-3, anti-HDAC-4, anti-HDAC-5, anti-HDAC-6 (all 1:100, Abcam, Cambridge, UK), or normal immunoglobulin G (IgG) as negative control; this was performed overnight at 4 °C.

    Techniques: Expressing, Comparison, Negative Control

    Immunohistochemical expression of HDAC-2 ( A , D ), HDAC-4 ( B , E ), and HDAC-5 ( C , F ) in epithelial ovarian carcinomas. ( A ) A high-grade serous ovarian carcinoma with nuclear HDAC-2 expression (×200). ( B ) A high-grade serous ovarian carcinoma with cytoplasmic HDAC-4 expression (×200). ( C ) A high-grade serous ovarian carcinoma with cytoplasmic HDAC-5 expression (×400). ( D ) A high-grade serous ovarian carcinoma with nuclear HDAC-2 expression (×400). ( E ) A high-grade serous ovarian carcinoma with cytoplasmic HDAC-4 expression (×400). ( F ) A high-grade serous ovarian carcinoma with nuclear and cytoplasmic HDAC-4 expression (×400).

    Journal: Biomedicines

    Article Title: Clinical Significance of the Immunohistochemical Expression of Histone Deacetylases (HDACs)-2, -4, and -5 in Ovarian Adenocarcinomas

    doi: 10.3390/biomedicines12050947

    Figure Lengend Snippet: Immunohistochemical expression of HDAC-2 ( A , D ), HDAC-4 ( B , E ), and HDAC-5 ( C , F ) in epithelial ovarian carcinomas. ( A ) A high-grade serous ovarian carcinoma with nuclear HDAC-2 expression (×200). ( B ) A high-grade serous ovarian carcinoma with cytoplasmic HDAC-4 expression (×200). ( C ) A high-grade serous ovarian carcinoma with cytoplasmic HDAC-5 expression (×400). ( D ) A high-grade serous ovarian carcinoma with nuclear HDAC-2 expression (×400). ( E ) A high-grade serous ovarian carcinoma with cytoplasmic HDAC-4 expression (×400). ( F ) A high-grade serous ovarian carcinoma with nuclear and cytoplasmic HDAC-4 expression (×400).

    Article Snippet: Subsequently, sections were stained with hematoxylin and eosin using an Automatic Stainer (BenchMark Ultra, VentanaRoche; Roche Diagnostics GmbH, Germany), according to the manufacturer’s protocol and using the following antibodies: rabbit polyclonal anti-HDAC-2 (Y461, ab32117, Abcam, Cambridge, U.K.), mouse monoclonal anti-HDAC-4 (ab16339, Abcam, Cambridge, UK), and mouse monoclonal anti-HDAC-5 (ab55403, Abcam, Cambridge, UK).

    Techniques: Immunohistochemical staining, Expressing

    Associations between HDAC-2 nuclear expression and serous histology ( first panel ) and with the presence of lymph node metastasis at the time of diagnosis in serous carcinomas ( second panel ).

    Journal: Biomedicines

    Article Title: Clinical Significance of the Immunohistochemical Expression of Histone Deacetylases (HDACs)-2, -4, and -5 in Ovarian Adenocarcinomas

    doi: 10.3390/biomedicines12050947

    Figure Lengend Snippet: Associations between HDAC-2 nuclear expression and serous histology ( first panel ) and with the presence of lymph node metastasis at the time of diagnosis in serous carcinomas ( second panel ).

    Article Snippet: Subsequently, sections were stained with hematoxylin and eosin using an Automatic Stainer (BenchMark Ultra, VentanaRoche; Roche Diagnostics GmbH, Germany), according to the manufacturer’s protocol and using the following antibodies: rabbit polyclonal anti-HDAC-2 (Y461, ab32117, Abcam, Cambridge, U.K.), mouse monoclonal anti-HDAC-4 (ab16339, Abcam, Cambridge, UK), and mouse monoclonal anti-HDAC-5 (ab55403, Abcam, Cambridge, UK).

    Techniques: Expressing