hdac3  (Proteintech)

Journal: Cell & Bioscience

Article Title: Microglial HDAC3 inhibits the migration of CD8-positive T cell in the development of EAE by restraining the expression of CCL5

doi: 10.1186/s13578-026-01564-5

Figure Lengend Snippet: HDAC3 is upregulated in microglia of the spinal cord from EAE model mice. A , B The protein levels of HDAC3 and GAPDH in the spinal cord from ctrl ( n = 4) and EAE ( n = 4) mice were determined by western blot ( A ) and the gray values were analyzed by Image J ( B ). C , D The mRNA ( n = 6 for each group) and protein ( n = 3 for each group) levels of HDAC3 in primary microglia isolated from Ctrl and EAE mice were determined by real-time PCR ( C ) and western blot ( D ). E , F The protein levels of HDAC3 in Iba1 positive cells in the spinal cord from ctrl and EAE mice were determined by immunostaining ( E ) and the fluorescence intensity were quantified by Image J ( F ). G – I The protein levels of HDAC3 in CD11b + CD45 low cells in the spinal cord from ctrl ( n = 4) and EAE ( n = 4) mice were determined by flow cytometry ( G , H ) and the mean fluorescent intensity of HDAC3 in CD11b + CD45 low cells were analyzed ( I ) (*indicates p < 0.05, **indicates p < 0.01 by Student’s t -test)

Article Snippet: The Hdac3 flox/+ mice (Stock No: 024119) and Cx3cr1 creERT2−IRES−EYFP (also named Cx3cr1 creERT2 ) transgenic mice (Stock No: 021160) were purchased from the Jackson Laboratory (Sacramento, CA, USA) and have been used in our previous study [ ].

Techniques: Western Blot, Isolation, Real-time Polymerase Chain Reaction, Immunostaining, Fluorescence, Flow Cytometry

Journal: Cell & Bioscience

Article Title: Microglial HDAC3 inhibits the migration of CD8-positive T cell in the development of EAE by restraining the expression of CCL5

doi: 10.1186/s13578-026-01564-5

Figure Lengend Snippet: Mice with microglial Hdac3-deficiency exhibited higher sensitivity to EAE induction. A Spinal cord slices from WT and Hdac3 cKO mice were stained with anti-HDAC3 (red) and anti-Iba1 (green) antibody, and the yellow arrow pointed to the signal of HDAC3 responding to the position of the Iba1-positive area. B The protein levels of HDAC3 and GAPDH in the primary microglia isolated from WT ( n = 3) and Hdac3 cKO ( n = 3) mice were determined by western blot and the gray values were analyzed by Image J. C Clinical score of WT EAE ( n = 8) and Hdac3 cKO EAE ( n = 8) mice were recorded every day post-immunization. D , E Spinal cord slices from WT EAE ( n = 6) and Hdac3 cKO EAE ( n = 6) mice were stained with Fast-blue and eosin ( D ), and the percentage of the Fast-blue negative area ( E ) was analyzed by ImageJ. F Spinal cord slices from WT EAE and Hdac3 cKO EAE mice were stained with anti-MBP antibody. G , H Spinal cord slices from WT EAE ( n = 7) and Hdac3 cKO EAE ( n = 7) mice were stained with H/E staining ( G ), and the number of infiltrated cells was counted manually ( H ). (*indicates p < 0.05, **indicates p < 0.01 by ANOVA or Student’s t -test)

Article Snippet: The Hdac3 flox/+ mice (Stock No: 024119) and Cx3cr1 creERT2−IRES−EYFP (also named Cx3cr1 creERT2 ) transgenic mice (Stock No: 021160) were purchased from the Jackson Laboratory (Sacramento, CA, USA) and have been used in our previous study [ ].

Techniques: Staining, Isolation, Western Blot

Journal: Cell & Bioscience

Article Title: Microglial HDAC3 inhibits the migration of CD8-positive T cell in the development of EAE by restraining the expression of CCL5

doi: 10.1186/s13578-026-01564-5

Figure Lengend Snippet: Microglial Hdac3-deficiency promoted periphery immune infiltration in the spinal cord during EAE development. A , B Kyoto encyclopedia of genes and genomes (KEGG) analysis ( A ) and ImmunoSystemProcess analysis ( B ) of the different expressed genes (DEGs) between the spinal cord from WT EAE mice and Hdac3 cKO EAE mice in RNA-sequencing data were performed by Cytoscape. C The mRNA levels of marker genes for T cell, B cell, NK cell and granulocyte in RNA-sequencing data from WT EAE mice and Hdac3 cKO EAE mice were displayed as heatmap. D , E Gene set enrichment analysis (GSEA) showed that the TCR signaling pathway ( D ) and positive regulation of leukocyte migration ( E ) were upregulated in the spinal cord from Hdac3 cKO EAE mice. F , G Genes involved in the TCR signaling pathway ( F ) and positive regulation of leukocyte migration ( G ) were upregulated in the spinal cord from Hdac3 cKO EAE mice ( N = 3 for each group)

Article Snippet: The Hdac3 flox/+ mice (Stock No: 024119) and Cx3cr1 creERT2−IRES−EYFP (also named Cx3cr1 creERT2 ) transgenic mice (Stock No: 021160) were purchased from the Jackson Laboratory (Sacramento, CA, USA) and have been used in our previous study [ ].

Techniques: RNA Sequencing, Marker, Migration

Journal: Cell & Bioscience

Article Title: Microglial HDAC3 inhibits the migration of CD8-positive T cell in the development of EAE by restraining the expression of CCL5

doi: 10.1186/s13578-026-01564-5

Figure Lengend Snippet: Inhibition of HDAC3 increased CD8 + T cell infiltration in spinal cord during EAE development. A – C The mRNA levels of CD3g ( A ), CD8a ( B ) and FasL ( C ) in the spinal cord from WT Ctrl ( n = 4), EAE mice administrated with RGFP966 ( n = 5) or vehicle ( n = 5) were determined by real-time PCR. D – F The mRNA levels of CD3g ( D ), CD8a ( E ) and FasL ( F ) in the spinal cord from WT Ctrl ( n = 6), WT EAE ( n = 6) and Hdac3 cKO EAE ( n = 6) mice were determined by real-time PCR. G – I The number of CD8 + T cell in the spinal cord from WT Ctrl, EAE mice administered with RGFP966 ( n = 6) or vehicle ( n = 6) ( G , H ), and WT EAE ( n = 5) and Hdac3 cKO EAE ( n = 5) mice ( G and I ) were detected by immunostaining with anti-CD8a and the number of CD8a-positive cells were counted. J , K The percentage of CD8 + T cells (gated in CD8a and CD3e double positive) in the spinal cord from WT Ctrl ( n = 6), WT EAE ( n = 6) and Hdac3 cKO EAE ( n = 6) mice were analyzed by flow cytometry with antibodies against CD8a and CD3e. (*indicates p < 0.05, **indicates p < 0.01 by ANOVA or Student’s t -test)

Article Snippet: The Hdac3 flox/+ mice (Stock No: 024119) and Cx3cr1 creERT2−IRES−EYFP (also named Cx3cr1 creERT2 ) transgenic mice (Stock No: 021160) were purchased from the Jackson Laboratory (Sacramento, CA, USA) and have been used in our previous study [ ].

Techniques: Inhibition, Real-time Polymerase Chain Reaction, Immunostaining, Flow Cytometry

Journal: Cell & Bioscience

Article Title: Microglial HDAC3 inhibits the migration of CD8-positive T cell in the development of EAE by restraining the expression of CCL5

doi: 10.1186/s13578-026-01564-5

Figure Lengend Snippet: Inhibition of HDAC3 increased the expression of CCL5 in spinal cord during EAE development. A Scatter diagram of the fold_change and p _value of the mRNA levels of chemokines between WT and Hdac3 cKO primary microglia analyzed by RNA-sequencing. B The chemokines that upregulated in Hdac3 cKO EAE spinal cord and Hdac3 cKO primary microglia were analyzed by RNA-sequencing. C , D The mRNA levels of CCL5 in the spinal cord from Ctrl ( n = 4), EAE mice administered with RGFP966 ( n = 7) or vehicle ( n = 5) ( C ), and WT_EAE ( n = 6) and Hdac3 cKO_EAE ( n = 6) mice ( D ) were determined by real-time PCR. E , F The mRNA levels of CCL5 ( E ) and HDAC3 ( F ) in WT ( n = 3) and Hdac3 cKO ( n = 3) primary microglia were determined by real-time PCR. G – I The protein levels of CCL5 and Iba1 in the spinal cord from Ctrl, EAE mice administrated with RGFP966 ( n = 6) or vehicle ( n = 6) ( G , H ), and WT_EAE ( n = 6) and Hdac3 cKO_EAE ( n = 6) mice ( G and I ) were detected by immunostaining with anti-CCL5 and anti-Iba1 antibodies and the number of CCL5 + Iba1 + cells were counted. (*indicates p < 0.05, **indicates p < 0.01 by ANOVA or Student’s t -test)

Article Snippet: The Hdac3 flox/+ mice (Stock No: 024119) and Cx3cr1 creERT2−IRES−EYFP (also named Cx3cr1 creERT2 ) transgenic mice (Stock No: 021160) were purchased from the Jackson Laboratory (Sacramento, CA, USA) and have been used in our previous study [ ].

Techniques: Inhibition, Expressing, RNA Sequencing, Real-time Polymerase Chain Reaction, Immunostaining

Journal: Cell & Bioscience

Article Title: Microglial HDAC3 inhibits the migration of CD8-positive T cell in the development of EAE by restraining the expression of CCL5

doi: 10.1186/s13578-026-01564-5

Figure Lengend Snippet: Microglial HDAC3 restrains IFN-γ induced expression of CCL5 by deacetylating on histone 3 lysine 9. A , B The mRNA levels ( A ) and protein levels ( B ) of HDAC3 and β-actin in N9 cells transfected with siRNA against HDAC3 (siHDAC3) or negative control (siNC) were determined by real-time PCR 72 h post-transfection. C , D N9 cells were pretreated with siRNA ( C ) or RGFP966 ( D ) as above, and then were stimulated with rIFN-γ (10 ng/ml) for indicating hours, and the cells were collected for detecting the mRNA levels of CCL5 by real-time PCR. E , F N9 cells were pretreated with siRNA ( E ) or RGFP966 ( F ) as above, and then were stimulated with rIFN-γ (10 ng/ml) for 24 h, and the protein levels of CCL5 in the supernatants were determined by ELISA. G N9 cells were pretreated with DMSO or RGFP966 (10 µM) for 12 h, then the cells were stimulated with rIFN-γ for indicating hours, and the protein levels of CCL5 in the supernatant and the protein levels of GAPDH in cells were determined via western blot. H Primary microglia were pretreated with RGFP966 for 12 h, and then were stimulated with rIFN-γ for 24 h, and the concentrations of CCL5 in the supernatant were determined by ELISA. I - J Migrated CD8 + T cells induced by conditional medium from primary microglia treated with rIFN-γ plus siRNA ( I ) or rIFN-γ plus RGFP966 ( J ) were analyzed by cell counting. K , L Migrated CD8 + T cells induced by conditional medium from N9 cells treated with rIFN-γ plus siRNA ( K ) or rIFN-γ plus RGFP966 ( L ) with or without CCL5 neutralization antibody were analyzed by cell counting. M The acetylation levels at H3K9 on the promoter of Ccl5 were determined by chromosome immunoprecipitation plus real-time PCR. M The model that HDAC3 regulates the expression of CCL5 by histone deacetylation. (* indicates p < 0.05, **indicates p < 0.01 by ANOVA or Student’s t -test)

Article Snippet: The Hdac3 flox/+ mice (Stock No: 024119) and Cx3cr1 creERT2−IRES−EYFP (also named Cx3cr1 creERT2 ) transgenic mice (Stock No: 021160) were purchased from the Jackson Laboratory (Sacramento, CA, USA) and have been used in our previous study [ ].

Techniques: Expressing, Transfection, Negative Control, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot, Cell Counting, Neutralization, Immunoprecipitation

Journal: Cell & Bioscience

Article Title: Microglial HDAC3 inhibits the migration of CD8-positive T cell in the development of EAE by restraining the expression of CCL5

doi: 10.1186/s13578-026-01564-5

Figure Lengend Snippet: Anti-CD8 neutralizing antibody could attenuate the symptoms of Hdac3 cKO EAE mice. A Clinical score of Hdac3 cKO_EAE mice injected with anti-CD8 NAb ( n = 7) or isotype IgG ( n = 7) every other day, and WT_EAE ( n = 7) mice injected with isotype IgG were recorded every day post immunization. B , C Spinal cord slices from Hdac3 cKO_EAE mice injected with anti-CD8 NAb ( n = 6) or isotype IgG ( n = 6) every other day, and WT_EAE ( n = 6) mice injected with isotype IgG were stained with Fast-blue and eosin ( B ), and the percentage of the Fast-blue negative area ( C ) was analyzed by ImageJ. D , E Spinal cord slices from Hdac3 cKO_EAE mice injected with anti-CD8 NAb ( n = 6) or isotype IgG ( n = 6) every other day, and WT_EAE ( n = 7) mice injected with isotype IgG were stained with anti-MBP antibody and the percentage of the Fast-blue negative area was analyzed by ImageJ. F Proposed mechanism: microglial HDAC3 restrained IFN-γ-induced expression of CCL5 via deacetylation of H3K9 on the promoter of Ccl5 ; inhibition of HDAC3 resulted in upregulation of CCL5 in microglia, which promoted the migration of CD8 + T cells to the spinal cord to accelerate the development of EAE

Article Snippet: The Hdac3 flox/+ mice (Stock No: 024119) and Cx3cr1 creERT2−IRES−EYFP (also named Cx3cr1 creERT2 ) transgenic mice (Stock No: 021160) were purchased from the Jackson Laboratory (Sacramento, CA, USA) and have been used in our previous study [ ].

Techniques: Injection, Staining, Expressing, Inhibition, Migration

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: Representative images of retina flatmount immunolabeling ( A ) at 14 days post-injury and quantitative analyses ( B ) demonstrate decreased neurodegeneration indicated by the neuronal marker NeuN and a reduction in microglia/ macrophage numbers, marked by Iba-1 ( C ) in the M-HDAC3 −/− retinas ( N = 5) compared to control HDAC3 f/f retinas ( N = 7). FOV = Field of view, * p < 0.05, *** p < 0.005, **** p < 0.001.

Article Snippet: Sections were then labeled for Iba-1, CD68 (BioLegend, Cat. #137002), MerTk (R&D Systems, Cat. # AF591), MBP (Abcam, Cat. # ab7349), HDAC3 (Proteintech, Cat. # 10255-1-AP), and GFAP (Abcam, Cat. #13-0300).

Techniques: Immunolabeling, Marker, Control

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: A , B Representative N1, P1, and N2 waveforms in the retinas of HDAC3 f/f and M-HDAC3 −/− sham and injured mice, conducted on day 7 post-ONC injury. Quantification and comparison of the ONC groups reveal improved waveform amplitudes in M-HDAC3 −/− retinas with statistical significance achieved in N2 and P1-N2 amplitudes compared to HDAC3 f/f retinas at 7 days post-ONC injury (HDAC3 f/f , N = 6 ; M-HDAC3 −/− , N = 7 )( C – F ), with no effect on the wave latencies ( G – I ). Similarly, N2 and P1-N2 amplitudes were significantly improved ( J – N ), but not their latencies ( O – Q ) at 14 days post-ONC injury (HDAC3 f/f , N = 5; M-HDAC3 −/− , N = 6). * p < 0.05, ** p < 0.01, **** p < 0.001, ns not statistically significant.

Article Snippet: Sections were then labeled for Iba-1, CD68 (BioLegend, Cat. #137002), MerTk (R&D Systems, Cat. # AF591), MBP (Abcam, Cat. # ab7349), HDAC3 (Proteintech, Cat. # 10255-1-AP), and GFAP (Abcam, Cat. #13-0300).

Techniques: Comparison

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: M-HDAC3 −/− and HDAC3 f/f mice were subjected to ONC, and retinas were collected at days 5 (HDAC3 f/f , N = 8; M-HDAC3 −/− , N = 15), 7, and 14 days (HDAC3 f/f , N = 4; M-HDAC3 −/− , N = 5) post-injury. A Representative Z-Stack confocal images of retina flatmounts at day 5 post-ONC injury display colocalization of TUNEL + apoptotic cells (red) and Iba-1 + microglia/macrophages (green). Arrows indicate free TUNEL + apoptotic cells, while arrowheads denote Iba-1-associated apoptotic cells. B Magnification of the crosshair area from the orthogonal view and 3D rendering showing a myeloid cell wrapping its processes around an apoptotic cell. C The ratio of engulfed apoptotic cells by microglia/macrophages (Iba-1 + TUNEL + ) to free apoptotic cells was markedly increased in injured retinas of M-HDAC3 −/− mice compared to HDAC3 f/f mice, indicating improved efferocytosis on day 5 after ONC. * p < 0.05.

Article Snippet: Sections were then labeled for Iba-1, CD68 (BioLegend, Cat. #137002), MerTk (R&D Systems, Cat. # AF591), MBP (Abcam, Cat. # ab7349), HDAC3 (Proteintech, Cat. # 10255-1-AP), and GFAP (Abcam, Cat. #13-0300).

Techniques: TUNEL Assay

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: A Representative confocal images of optic nerve sections from M-HDAC3 −/− and HDAC3 f/f mice immunolabeled with Iba-1 (myeloid cell marker, green), CD68 (phagocytic cell marker, red), and DAPI (nuclei marker, blue) demonstrate an increase in phagocytic myeloid cells, indicated by arrows, in M-HDAC3 −/− mice compared to HDAC3 f/f on day 7 after ONC. B Representative confocal images of axonal growth and nerve fiber sprouting in the axons distal to the crush site by anterograde tracing with cholera toxin B (CTB) on day 14 post-ONC. C The quantification of the sprouting axons demonstrated significant improvement in axonal growth in M-HDAC3 −/− compared to HDAC3 f/f retinas, indicated by fluorescence intensity at distances of 200, 400, and 600 μm beyond the crush site (HDAC3 f/f , N = 5; M-HDAC3 −/− , N = 7). * p < 0.05, ** p < 0.01, *** p < 0.005.

Article Snippet: Sections were then labeled for Iba-1, CD68 (BioLegend, Cat. #137002), MerTk (R&D Systems, Cat. # AF591), MBP (Abcam, Cat. # ab7349), HDAC3 (Proteintech, Cat. # 10255-1-AP), and GFAP (Abcam, Cat. #13-0300).

Techniques: Immunolabeling, Marker, Anterograde Tracing, Fluorescence

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: A Experimental setup using myelin debris from the optic nerve labeled with Dil-red dye (top) and unlabeled debris stained with Oil Red O (ORO, bottom). B Representative images show the internalization of DiI-labeled myelin (arrows) by bone-marrow-derived macrophages derived from HDAC3 f/f and M-HDAC3 −/− mice. C Quantification of uptake of DiI-labeled myelin, expressed as mean fluorescence intensity (MFI), demonstrates significant improvement in the phagocytic activity of M-HDAC3 −/− macrophages compared to HDAC3 f/f macrophages. D ORO staining confirmed the improved uptake of myelin debris (arrows) by M-HDAC3 −/− macrophages compared to HDAC3 f/f macrophages. E Representative confocal images of Iba-1+ myeloid cells (red) and myelin basic protein (MBP, green) in optic nerve sections 7 days after ONC show a considerable increase in myelin clearance by myeloid cells in M-HDAC3 −/− optic nerves compared to HDAC3 f/f injured controls, as evidenced by increased Iba-1/MBP colocalization. N = 3 per group, **** p < 0.001.

Article Snippet: Sections were then labeled for Iba-1, CD68 (BioLegend, Cat. #137002), MerTk (R&D Systems, Cat. # AF591), MBP (Abcam, Cat. # ab7349), HDAC3 (Proteintech, Cat. # 10255-1-AP), and GFAP (Abcam, Cat. #13-0300).

Techniques: Labeling, Staining, Derivative Assay, Fluorescence, Activity Assay

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: Macrophages from HDAC3 f/f and M-HDAC3 −/− mice were co-incubated with K-562 apoptotic cells (apop) in an in vitro efferocytosis assay. Controls included either co-incubation of macrophages with K-562 non-apoptotic cells (non-apop) or no treatment (no ttt). A – C Quantification of mRNA levels of ODC, MerTK, and the anti-inflammatory cytokine IL-10 demonstrated significant upregulation in M-HDAC3 −/− macrophages as compared to HDAC3 f/f macrophages incubated with apoptotic cells. D – F Western blotting shows significant upregulation of MerTK and ODC in M-HDAC3 −/− macrophages co-cultured with K-562 cells compared to untreated M-HDAC3 −/− macrophages, but not in the HDAC3 f/f control co-cultures. β-actin was used as a loading control. G Representative confocal images of Iba-1 + myeloid cells (green) and MerTK (red) in retinal sections 7 days after ONC show a considerable increase in MerTK expression by myeloid cells in M-HDAC3 −/− retinas compared to HDAC3 f/f controls, as evidenced by increased Iba-1/MerTK colocalization. H Similarly, MerTK expression by myeloid cells is increased in injured optic nerve sections, with arrowheads pointing to Iba-1 + MerTK + myeloid cells. GCl ganglion cell layer, INL inner nuclear layer, ONL outer nuclear layer. N = 4 per group. * p < 0.05, *** p < 0.005, **** p < 0.001.

Article Snippet: Sections were then labeled for Iba-1, CD68 (BioLegend, Cat. #137002), MerTk (R&D Systems, Cat. # AF591), MBP (Abcam, Cat. # ab7349), HDAC3 (Proteintech, Cat. # 10255-1-AP), and GFAP (Abcam, Cat. #13-0300).

Techniques: Incubation, In Vitro, Western Blot, Cell Culture, Control, Expressing

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: A Representative images illustrate the internalization of DiI-labeled myelin debris (red), derived from optic nerve axons, by CFDA-labeled macrophages (green) obtained from HDAC3 f/f and M-HDAC3 −/− mice ( N = 5 per group). These cells were pretreated with 0.8 nM of the MertK inhibitor (UNC2025) or vehicle for 1 h in vitro. B Quantification of DiI-labeled myelin debris uptake, expressed as mean fluorescence intensity (MFI), demonstrates a significant reduction in myelin uptake in the UNC2025 pretreatment groups, with UNC2025 abolishing the enhanced myelin uptake observed in vehicle-treated M-HDAC3 −/− macrophages, indicating that myeloid HDAC3 deletion promotes myelin uptake at least in part via MerTK. C Representative images of immunolabeling for neurons, marked by NeuN (green), and microglia/macrophages, marked by Iba-1 (red), of adult M-HDAC3 −/− and HDAC3 f/f mice retinas that were explanted for 24 h and treated with UNC2025 or vehicle (HDAC3 f/f , N = 3, 4; M-HDAC3 −/− , N = 5) for another 24 h. D , E Quantification of NeuN and Iba-1 shows that UNC2025 treatments had no significant effect on neuronal preservation in retinal explants of both treated groups, while it significantly increased myeloid cell number in M-HDAC3 −/− compared to flox retina explants. F , G Representative images of Iba-1 (red) labeling and quantification of optic nerve explants treated with UNC2025 show no effect of the treatment on myeloid cell count between M-HDAC3 −/− and HDAC3 f/f derived optic nerves ( N = 3 per group). * p < 0.05, **** p < 0.001; ns, not statistically significant; FOV, field of view.

Article Snippet: Sections were then labeled for Iba-1, CD68 (BioLegend, Cat. #137002), MerTk (R&D Systems, Cat. # AF591), MBP (Abcam, Cat. # ab7349), HDAC3 (Proteintech, Cat. # 10255-1-AP), and GFAP (Abcam, Cat. #13-0300).

Techniques: Labeling, Derivative Assay, In Vitro, Fluorescence, Immunolabeling, Preserving, Cell Characterization

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: A Representative retina flatmount images from microglia-specific HDAC3 KO (im-HDAC3 −/− ) and HDAC3 f/f controls immunolabeled for NeuN (neuronal marker, green) and Iba-1 (microglia/macrophages marker, red) at 14 days after ONC. B , C Quantitative analyses reveal significant neurodegeneration, indicated by a decrease in the neuronal marker NeuN and increase in Iba-1 + cell count in injured im-HDAC3 −/− ( N = 5) and HDAC3 f/f ( N = 6) mice compared to shams ( N = 3 for im-HDAC3 −/− , and N = 4 for HDAC3 f/f ). However, no differences were observed between the injured groups. D , E Representative images and Iba-1 quantification at the optic nerve injury site of im-HDAC3 −/− ( N = 5) and HDAC3 f/f ( N = 4) mice 14 days post-ONC injury show a robust presence of activated microglia and macrophages, with no differences observed between the injured groups. *** p < 0.005, **** p < 0.001; ns, not statistically significant; FOV, field of view.

Article Snippet: Sections were then labeled for Iba-1, CD68 (BioLegend, Cat. #137002), MerTk (R&D Systems, Cat. # AF591), MBP (Abcam, Cat. # ab7349), HDAC3 (Proteintech, Cat. # 10255-1-AP), and GFAP (Abcam, Cat. #13-0300).

Techniques: Immunolabeling, Marker, Cell Characterization

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: Representative images of retina flatmount immunolabeling ( A ) at 14 days post-injury and quantitative analyses ( B ) demonstrate decreased neurodegeneration indicated by the neuronal marker NeuN and a reduction in microglia/ macrophage numbers, marked by Iba-1 ( C ) in the M-HDAC3 −/− retinas ( N = 5) compared to control HDAC3 f/f retinas ( N = 7). FOV = Field of view, * p < 0.05, *** p < 0.005, **** p < 0.001.

Article Snippet: Two myeloid HDAC3 KO lines were used (genotyping is included in Supplementary Fig. ): A constitutive macrophage and microglia cell-specific HDAC3 KO (LysM Cre/+ ; HDAC3 f/f or M-HDAC3 −/− ) mice generated by crossing the HDAC3 f/f mice with LysM Cre/+ mice (Jackson laboratory Stock # 004781).

Techniques: Immunolabeling, Marker, Control

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: A , B Representative N1, P1, and N2 waveforms in the retinas of HDAC3 f/f and M-HDAC3 −/− sham and injured mice, conducted on day 7 post-ONC injury. Quantification and comparison of the ONC groups reveal improved waveform amplitudes in M-HDAC3 −/− retinas with statistical significance achieved in N2 and P1-N2 amplitudes compared to HDAC3 f/f retinas at 7 days post-ONC injury (HDAC3 f/f , N = 6 ; M-HDAC3 −/− , N = 7 )( C – F ), with no effect on the wave latencies ( G – I ). Similarly, N2 and P1-N2 amplitudes were significantly improved ( J – N ), but not their latencies ( O – Q ) at 14 days post-ONC injury (HDAC3 f/f , N = 5; M-HDAC3 −/− , N = 6). * p < 0.05, ** p < 0.01, **** p < 0.001, ns not statistically significant.

Article Snippet: Two myeloid HDAC3 KO lines were used (genotyping is included in Supplementary Fig. ): A constitutive macrophage and microglia cell-specific HDAC3 KO (LysM Cre/+ ; HDAC3 f/f or M-HDAC3 −/− ) mice generated by crossing the HDAC3 f/f mice with LysM Cre/+ mice (Jackson laboratory Stock # 004781).

Techniques: Comparison

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: M-HDAC3 −/− and HDAC3 f/f mice were subjected to ONC, and retinas were collected at days 5 (HDAC3 f/f , N = 8; M-HDAC3 −/− , N = 15), 7, and 14 days (HDAC3 f/f , N = 4; M-HDAC3 −/− , N = 5) post-injury. A Representative Z-Stack confocal images of retina flatmounts at day 5 post-ONC injury display colocalization of TUNEL + apoptotic cells (red) and Iba-1 + microglia/macrophages (green). Arrows indicate free TUNEL + apoptotic cells, while arrowheads denote Iba-1-associated apoptotic cells. B Magnification of the crosshair area from the orthogonal view and 3D rendering showing a myeloid cell wrapping its processes around an apoptotic cell. C The ratio of engulfed apoptotic cells by microglia/macrophages (Iba-1 + TUNEL + ) to free apoptotic cells was markedly increased in injured retinas of M-HDAC3 −/− mice compared to HDAC3 f/f mice, indicating improved efferocytosis on day 5 after ONC. * p < 0.05.

Article Snippet: Two myeloid HDAC3 KO lines were used (genotyping is included in Supplementary Fig. ): A constitutive macrophage and microglia cell-specific HDAC3 KO (LysM Cre/+ ; HDAC3 f/f or M-HDAC3 −/− ) mice generated by crossing the HDAC3 f/f mice with LysM Cre/+ mice (Jackson laboratory Stock # 004781).

Techniques: TUNEL Assay

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: A Representative confocal images of optic nerve sections from M-HDAC3 −/− and HDAC3 f/f mice immunolabeled with Iba-1 (myeloid cell marker, green), CD68 (phagocytic cell marker, red), and DAPI (nuclei marker, blue) demonstrate an increase in phagocytic myeloid cells, indicated by arrows, in M-HDAC3 −/− mice compared to HDAC3 f/f on day 7 after ONC. B Representative confocal images of axonal growth and nerve fiber sprouting in the axons distal to the crush site by anterograde tracing with cholera toxin B (CTB) on day 14 post-ONC. C The quantification of the sprouting axons demonstrated significant improvement in axonal growth in M-HDAC3 −/− compared to HDAC3 f/f retinas, indicated by fluorescence intensity at distances of 200, 400, and 600 μm beyond the crush site (HDAC3 f/f , N = 5; M-HDAC3 −/− , N = 7). * p < 0.05, ** p < 0.01, *** p < 0.005.

Article Snippet: Two myeloid HDAC3 KO lines were used (genotyping is included in Supplementary Fig. ): A constitutive macrophage and microglia cell-specific HDAC3 KO (LysM Cre/+ ; HDAC3 f/f or M-HDAC3 −/− ) mice generated by crossing the HDAC3 f/f mice with LysM Cre/+ mice (Jackson laboratory Stock # 004781).

Techniques: Immunolabeling, Marker, Anterograde Tracing, Fluorescence

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: A Experimental setup using myelin debris from the optic nerve labeled with Dil-red dye (top) and unlabeled debris stained with Oil Red O (ORO, bottom). B Representative images show the internalization of DiI-labeled myelin (arrows) by bone-marrow-derived macrophages derived from HDAC3 f/f and M-HDAC3 −/− mice. C Quantification of uptake of DiI-labeled myelin, expressed as mean fluorescence intensity (MFI), demonstrates significant improvement in the phagocytic activity of M-HDAC3 −/− macrophages compared to HDAC3 f/f macrophages. D ORO staining confirmed the improved uptake of myelin debris (arrows) by M-HDAC3 −/− macrophages compared to HDAC3 f/f macrophages. E Representative confocal images of Iba-1+ myeloid cells (red) and myelin basic protein (MBP, green) in optic nerve sections 7 days after ONC show a considerable increase in myelin clearance by myeloid cells in M-HDAC3 −/− optic nerves compared to HDAC3 f/f injured controls, as evidenced by increased Iba-1/MBP colocalization. N = 3 per group, **** p < 0.001.

Article Snippet: Two myeloid HDAC3 KO lines were used (genotyping is included in Supplementary Fig. ): A constitutive macrophage and microglia cell-specific HDAC3 KO (LysM Cre/+ ; HDAC3 f/f or M-HDAC3 −/− ) mice generated by crossing the HDAC3 f/f mice with LysM Cre/+ mice (Jackson laboratory Stock # 004781).

Techniques: Labeling, Staining, Derivative Assay, Fluorescence, Activity Assay

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: Macrophages from HDAC3 f/f and M-HDAC3 −/− mice were co-incubated with K-562 apoptotic cells (apop) in an in vitro efferocytosis assay. Controls included either co-incubation of macrophages with K-562 non-apoptotic cells (non-apop) or no treatment (no ttt). A – C Quantification of mRNA levels of ODC, MerTK, and the anti-inflammatory cytokine IL-10 demonstrated significant upregulation in M-HDAC3 −/− macrophages as compared to HDAC3 f/f macrophages incubated with apoptotic cells. D – F Western blotting shows significant upregulation of MerTK and ODC in M-HDAC3 −/− macrophages co-cultured with K-562 cells compared to untreated M-HDAC3 −/− macrophages, but not in the HDAC3 f/f control co-cultures. β-actin was used as a loading control. G Representative confocal images of Iba-1 + myeloid cells (green) and MerTK (red) in retinal sections 7 days after ONC show a considerable increase in MerTK expression by myeloid cells in M-HDAC3 −/− retinas compared to HDAC3 f/f controls, as evidenced by increased Iba-1/MerTK colocalization. H Similarly, MerTK expression by myeloid cells is increased in injured optic nerve sections, with arrowheads pointing to Iba-1 + MerTK + myeloid cells. GCl ganglion cell layer, INL inner nuclear layer, ONL outer nuclear layer. N = 4 per group. * p < 0.05, *** p < 0.005, **** p < 0.001.

Article Snippet: Two myeloid HDAC3 KO lines were used (genotyping is included in Supplementary Fig. ): A constitutive macrophage and microglia cell-specific HDAC3 KO (LysM Cre/+ ; HDAC3 f/f or M-HDAC3 −/− ) mice generated by crossing the HDAC3 f/f mice with LysM Cre/+ mice (Jackson laboratory Stock # 004781).

Techniques: Incubation, In Vitro, Western Blot, Cell Culture, Control, Expressing

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: A Representative images illustrate the internalization of DiI-labeled myelin debris (red), derived from optic nerve axons, by CFDA-labeled macrophages (green) obtained from HDAC3 f/f and M-HDAC3 −/− mice ( N = 5 per group). These cells were pretreated with 0.8 nM of the MertK inhibitor (UNC2025) or vehicle for 1 h in vitro. B Quantification of DiI-labeled myelin debris uptake, expressed as mean fluorescence intensity (MFI), demonstrates a significant reduction in myelin uptake in the UNC2025 pretreatment groups, with UNC2025 abolishing the enhanced myelin uptake observed in vehicle-treated M-HDAC3 −/− macrophages, indicating that myeloid HDAC3 deletion promotes myelin uptake at least in part via MerTK. C Representative images of immunolabeling for neurons, marked by NeuN (green), and microglia/macrophages, marked by Iba-1 (red), of adult M-HDAC3 −/− and HDAC3 f/f mice retinas that were explanted for 24 h and treated with UNC2025 or vehicle (HDAC3 f/f , N = 3, 4; M-HDAC3 −/− , N = 5) for another 24 h. D , E Quantification of NeuN and Iba-1 shows that UNC2025 treatments had no significant effect on neuronal preservation in retinal explants of both treated groups, while it significantly increased myeloid cell number in M-HDAC3 −/− compared to flox retina explants. F , G Representative images of Iba-1 (red) labeling and quantification of optic nerve explants treated with UNC2025 show no effect of the treatment on myeloid cell count between M-HDAC3 −/− and HDAC3 f/f derived optic nerves ( N = 3 per group). * p < 0.05, **** p < 0.001; ns, not statistically significant; FOV, field of view.

Article Snippet: Two myeloid HDAC3 KO lines were used (genotyping is included in Supplementary Fig. ): A constitutive macrophage and microglia cell-specific HDAC3 KO (LysM Cre/+ ; HDAC3 f/f or M-HDAC3 −/− ) mice generated by crossing the HDAC3 f/f mice with LysM Cre/+ mice (Jackson laboratory Stock # 004781).

Techniques: Labeling, Derivative Assay, In Vitro, Fluorescence, Immunolabeling, Preserving, Cell Characterization

Journal: Cell Death Discovery

Article Title: Myeloid HDAC3 deletion protects against traumatic optic injury

doi: 10.1038/s41420-026-03030-0

Figure Lengend Snippet: A Representative retina flatmount images from microglia-specific HDAC3 KO (im-HDAC3 −/− ) and HDAC3 f/f controls immunolabeled for NeuN (neuronal marker, green) and Iba-1 (microglia/macrophages marker, red) at 14 days after ONC. B , C Quantitative analyses reveal significant neurodegeneration, indicated by a decrease in the neuronal marker NeuN and increase in Iba-1 + cell count in injured im-HDAC3 −/− ( N = 5) and HDAC3 f/f ( N = 6) mice compared to shams ( N = 3 for im-HDAC3 −/− , and N = 4 for HDAC3 f/f ). However, no differences were observed between the injured groups. D , E Representative images and Iba-1 quantification at the optic nerve injury site of im-HDAC3 −/− ( N = 5) and HDAC3 f/f ( N = 4) mice 14 days post-ONC injury show a robust presence of activated microglia and macrophages, with no differences observed between the injured groups. *** p < 0.005, **** p < 0.001; ns, not statistically significant; FOV, field of view.

Article Snippet: Two myeloid HDAC3 KO lines were used (genotyping is included in Supplementary Fig. ): A constitutive macrophage and microglia cell-specific HDAC3 KO (LysM Cre/+ ; HDAC3 f/f or M-HDAC3 −/− ) mice generated by crossing the HDAC3 f/f mice with LysM Cre/+ mice (Jackson laboratory Stock # 004781).

Techniques: Immunolabeling, Marker, Cell Characterization

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: HDAC3 Preserves the primordial follicle reserve by epigenetically suppressing ferroptosis in pregranulosa cells

doi: 10.1007/s00018-026-06128-x

Figure Lengend Snippet: HDAC3 played an important role in PFs maintenance. a , b HDAC3 (green) kept a stable expression level in pGCs before puberty and in adulthood. Nuclei were dyed with Hoechst (blue). PFs were framed by the yellow dashed line, pGCs were framed by white dashed line. Scale bar = 40 μm. n = 3 biologically independent experiments. a Germ cells were dyed with MVH (rad). b GCs were dyed with FOXL2 (red). c , e Pan-acetylation levels of ovarian proteins were significantly upregulated after the inhibition of HDAC3 in vitro. RGFP and SC were the two specific inhibitors of HDAC3. n = 3. Data are presented as mean ± SD. Asterisks (*) indicate significant differences at ** P <0.01. d Ovarian volume unchanged after inhibition of HDAC3 in 2 dpp ovaries cultured for 1 day, but ovarian volume was decreased after inhibition of HDAC3 in 2 dpp ovaries cultured for 5 days. Scale bar = 40 μm. n ≥ 3. f The number of PFs was decreased in a time-dependent manner by inhibiting HDAC3. n ≥ 3. Data are presented as mean ± SD. Asterisks (*) indicate significant differences at ** P <0.01. g The number of total follicles was decreased in a time-dependent manner by inhibiting HDAC3. n ≥ 3. Data are presented as mean ± SD. Asterisks (*) indicate significant differences at ** P <0.01

Article Snippet: Hdac3 flox/flox mice were purchased from the Jackson Laboratory (ME, USA), and Foxl2-CreER T2 mice were gifted by Prof. Kui Liu.

Techniques: Expressing, Inhibition, In Vitro, Cell Culture

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: HDAC3 Preserves the primordial follicle reserve by epigenetically suppressing ferroptosis in pregranulosa cells

doi: 10.1007/s00018-026-06128-x

Figure Lengend Snippet: Deficiency of Hdac3 in GCs resulted in decreased follicle number. a Schematic representation of the deletion of Hdac3 exon 4–7 in GCs was induced by tamoxifen injection at 1dpp, 3 dpp and 5 dpp. b HDAC3 levels in 10-dpp-old Hdac3 f/f and Hdac3 cKO ovaries were quantified by Western blotting analysis. n = 3 biologically independent experiments. c HDAC3 (green) were decreased in ovaries from 10-dpp-old Hdac3 cKO mice. Nuclei were dyed with Hoechst (blue). GCs were dyed with FOXL2 (red). Scale bar = 40 μm. n = 3 biologically independent experiments. d , e Comparison of follicle numbers of Hdac3 f/f and Hdac3 cKO mice at 10 dpp ( n = 4, p = 0.4972 for PFs, p = 0.8286 for PaFs, p = 0.3672 for SFs, p = 0.5039 for total follicles). Scale bar = 40 μm. f HDAC3 (green) were decreased in ovaries from 10-dpp-old Hdac3 cKO mice. Nuclei were dyed with Hoechst (blue). Scale bar = 40 μm. n = 3 biologically independent experiments. g , h Comparison of follicle numbers of Hdac3 f/f and Hdac3 cKO mice at 2 w ( n = 3, p = 0.0031 for PFs, p = 0.0134 for PaFs, p = 0.0002 for SFs, p = 0.0031 for total follicles). Scale bar = 40 μm. i HDAC3 (green) were decreased in ovaries from 2-week-old Hdac3 cKO mice. Nuclei were dyed with Hoechst (blue). Scale bar = 40 μm. n = 3. j , k Comparison of follicle numbers of Hdac3 f/f and Hdac3 cKO mice at 3 w ( n = 3, p = 0.0182 for PFs, p = 0.0222 for PaFs, p = 0.0242 for SFs, p = 0.0133 for total follicles). Scale bar = 40 μm. l HDAC3 (green) were decreased in ovaries from 3-week-old Hdac3 cKO mice. Nuclei were dyed with Hoechst (blue). Scale bar = 40 μm. n = 3

Article Snippet: Hdac3 flox/flox mice were purchased from the Jackson Laboratory (ME, USA), and Foxl2-CreER T2 mice were gifted by Prof. Kui Liu.

Techniques: Injection, Western Blot, Comparison

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: HDAC3 Preserves the primordial follicle reserve by epigenetically suppressing ferroptosis in pregranulosa cells

doi: 10.1007/s00018-026-06128-x

Figure Lengend Snippet: The inhibition of HDAC3 did not affect PF activation. a , b Representative images showed that the translocation of FOXO3a (green) from the nucleus to the cytoplasm (white arrows) was not facilitated after inhibition of HDAC3 by RGFP in 2 dpp ovaries cultured for 1 day. n = 5. Nuclei were dyed with Hoechst (blue). GCs were dyed with FOXL2 (rad). Data are presented as mean ± SD. Scale bar = 40 μm. c , d Representative images showed that the translocation of FOXO3a (green) from the nucleus to the cytoplasm (white arrows) was not facilitated after inhibiting HDAC3 (RGFP) in 2 dpp ovaries cultured for 2 days. n = 6 biologically independent experiments. Nuclei were dyed with Hoechst (blue). GCs were dyed with FOXL2 (rad). Data are presented as mean ± SD. Scale bar = 40 μm. e Western blotting analysis showed that the inhabitation of HDAC3 had no effects on the protein levels of molecules related to PF activation

Article Snippet: Hdac3 flox/flox mice were purchased from the Jackson Laboratory (ME, USA), and Foxl2-CreER T2 mice were gifted by Prof. Kui Liu.

Techniques: Inhibition, Activation Assay, Translocation Assay, Cell Culture, Western Blot

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: HDAC3 Preserves the primordial follicle reserve by epigenetically suppressing ferroptosis in pregranulosa cells

doi: 10.1007/s00018-026-06128-x

Figure Lengend Snippet: Inhibition of HDAC3 caused ferroptosis in cultured ovaries in vitro . a , b Lipid ROS in ovarian cells after RGFP treatment. Scale bar = 40 μm. n ≥ 3. Data are presented as mean ± SD. Asterisks (*) indicate significant differences at *** P < 0.001. c , d Representative images showing that ACSL4 (green) in control and HDAC3 inhibited ovaries. The level of ACSL4 was increased in ovaries after RGFP treatment, pGCs were framed by white dashed line. Scale bar = 40 μm. n = 3, *** P value<0.0001. e TEM of PFs in control and HDAC3 inhibited ovaries. Mitochondrial volume was decreased after the inhibiting of HDAC3 in 2 dpp ovaries cultured for 2 days. Scale bar = 40 μm. n = 3. f Western bolting assay showed that the expression levels of ferroptosis related proteins ACSL4 were increased after the inhibition of HDAC3 in 2 dpp ovaries cultured for 1 day. n = 3. g RNA-seq analysis revealed the transcriptional levels of Tfr2 were increased after HDAC3 inhibited (as indicated by the Log10(FPKM) of the RGFP group compared with that of the control group). n = 3, p value<0.001. h Ovarian volume was rescued after inhibiting of HDAC3 while inhibiting ferroptosis in 2 dpc ovaries cultured for 5 days. Scale bar = 40 μm. n ≥ 3. i The number of ovarian follicles was rescued by inhibiting HDAC3 while inhibition of ferroptosis in 2 dpc ovaries cultured for 5 days. n ≥ 3. Data are presented as mean ± SD. Asterisks (*) indicate significant differences at ** P <0.01. j Western bolting assay showed that the levels of ferroptosis related proteins ACSL4 were similar to control group after the inhibiting of HDAC3 while the inhibiting of ferroptosis in 2 dpp ovaries cultured for 5 days. n = 3. k , l Lipid ROS in ovarian cells after Erastin treatment. Scale bar = 40 μm. Data are presented as mean ± SD. n = 3, p value<0.0001. m Ovarian volume was decreased after activating of ferroptosis in 2 dpp ovaries cultured for 5 days. Scale bar = 40 μm. n ≥ 3. n The number of ovarian follicles was rescued by activating ferroptosis in 2 dpp ovaries cultured for 5 days. n ≥ 3. Data are presented as mean ± SD. Asterisks (*) indicate significant differences at ** P <0.01, *** P < 0.001

Article Snippet: Hdac3 flox/flox mice were purchased from the Jackson Laboratory (ME, USA), and Foxl2-CreER T2 mice were gifted by Prof. Kui Liu.

Techniques: Inhibition, Cell Culture, In Vitro, Control, Western Blot, Expressing, RNA Sequencing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: HDAC3 Preserves the primordial follicle reserve by epigenetically suppressing ferroptosis in pregranulosa cells

doi: 10.1007/s00018-026-06128-x

Figure Lengend Snippet: Inhibition of HDAC3 affected the expression of ferroptosis-related genes and iron homeostasis. a Heatmap showing the expression patterns of certain ferroptosis-related genes. b Total iron in GCs from the control and the HDAC3 inhibited ovaries (2 dpp + 1 day). c n(Fe 2+ )/n(Fe) in GCs from the control and the HDAC3 inhibited ovaries (2 dpp + 1 day). d Relative content of ferrous iron of cytoplasm and mitochondria in GCs from the control and the HDAC3 inhibited ovaries (2 dpp + 1 day). e-h Western blotting analysis of TFR2, FDXR, GOT1 and FUNDC2 in GCs from the control and the HDAC3 inhibited ovaries (2 dpp + 1 day). i Western blotting analysis of TF in the control and the HDAC3 inhibited ovaries (2 dpp + 1 day). j Western blot analysis to identify the separate effect of cytoplasmic and mitochondrial. k-o Statistical analysis of difference in protein levels of TFR2, FDXR, GOT1, FUNDC2 and TF. p-t Statistical analysis of difference in RNA levels of TFR2, FDXR, GOT1, FUNDC2 and TF. Total proteins and RNA values were compared to the control, which was normalized to 1.0. Data were presented as mean ± SEM, n ≥ 3. ‘’ns’’ indicates not significant ( P > 0.05), whereas the asterisks (*) indicate significant differences at ** P < 0.01, *** P < 0.001

Article Snippet: Hdac3 flox/flox mice were purchased from the Jackson Laboratory (ME, USA), and Foxl2-CreER T2 mice were gifted by Prof. Kui Liu.

Techniques: Inhibition, Expressing, Control, Western Blot

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: HDAC3 Preserves the primordial follicle reserve by epigenetically suppressing ferroptosis in pregranulosa cells

doi: 10.1007/s00018-026-06128-x

Figure Lengend Snippet: HDAC3 deficiency impaired levels of proteins involved in iron metabolism and ISCs in GCs. a-h Western blotting analysis of p53, p21, p16, IRP1, IRP2, FTMT, NDUFS1, SDHB and UQCRFS1 in GCs from 10-dpp-old Hdac3 f/f and Hdac3 cKO ovaries. i-q Statistical analysis of difference in protein levels of p53, p21, p16, IRP1, IRP2, FTMT, NDUFS1, SDHB and UQCRFS1. Total proteins values were compared to the control, which was normalized to 1.0. Data were presented as mean ± SEM, n ≥ 3. ‘’ns’’ indicates not significant ( P > 0.05), whereas the asterisks (*) indicate significant differences at * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Hdac3 flox/flox mice were purchased from the Jackson Laboratory (ME, USA), and Foxl2-CreER T2 mice were gifted by Prof. Kui Liu.

Techniques: Western Blot, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: HDAC3 Preserves the primordial follicle reserve by epigenetically suppressing ferroptosis in pregranulosa cells

doi: 10.1007/s00018-026-06128-x

Figure Lengend Snippet: H3K27ac affected the transcriptional activity of ferroptosis-related genes. a , c The expression level of H3K27ac was increased, but the expression level of H3K4ac, H3K9ac, H3K14ac, H3K18ac and H3K23ac were not changed in 2 dpp ovaries cultured with the inhibitor of HDAC3 for 1 day. n = 3. b The level of H3K27ac was increased, but the level of H3K9ac was not changed in Hdac3 cKO mice, n = 1. d The level of HDAC3 was decreased in Hdac3 cKO mice, n = 1. e Distribution of H3K27ac binding sites in the TSS region, with horizontal coordinates indicating the relative position of reads to the TSS and vertical coordinates indicating the coverage depth. f Distribution characteristics of H3K27ac binding sites. g Distribution of H3K27ac on ferroptosis-related genes and nearby cis-regulatory elements. h ChIP-qPCR analysis of the enrichment of H3K27ac in the promoter region of ferroptosis-related genes in GCs from 10-dpp-old C57B6/J mice. qPCR data were processed using the Livak method, and the values were normalised to show the multiplicity of enrichment. Data were presented as mean ± SEM, n ≥ 3. Asterisks (*) indicate significant differences at ** P < 0.01, *** P < 0.001

Article Snippet: Hdac3 flox/flox mice were purchased from the Jackson Laboratory (ME, USA), and Foxl2-CreER T2 mice were gifted by Prof. Kui Liu.

Techniques: Activity Assay, Expressing, Cell Culture, Binding Assay, ChIP-qPCR

Journal: Frontiers in Transplantation

Article Title: Mechanisms inducing differentiation of adult islet progenitor-like cells into functional islet-like organoids

doi: 10.3389/frtra.2026.1740314

Figure Lengend Snippet: Requirement of CN/NFAT signaling to recruit p300 to RFX6 and NEUROD1 promoters during ISX9-induced IPC islet organoid differentiation (A) induction of NFAT family isoform genes in IPC clusters treated with ISX9 for 24 h in the presence of CN inhibitor FK506. (B) RFX6 and NEUROD1 promoter activation by ISX9 in IPCs in the presence of FK506 or transfected with gene vectors overexpressing dominant negative NFAT (dnNFAT) and mutated control (dnNFATm). (C) ChIP assay of association of NFATC2, p300, HDAC1, HDAC2, and HDAC3 with RFX6, NEUROD1, and NEUROG3 promoters upon 6 h treatment of IPCs with ISX9 with and without 24 h pretreatment with ITF2357. Graphed values are expressed as mean ± SD. Asterisks above bars indicate statistically significant differences (* p < 0.05, *** p < 0.001) in mean values for treatments based on a two-way ANOVA and Sidak's multiple comparison test. Data shown are results from at least three independent experiments using IPCs derived from three individual donors.

Article Snippet: Antibodies used were as follows: NFATC2, RGS16, NEUROD1, NEUROG3, p300, HDAC1, HDAC2, and HDAC3 (Santa Cruz Biotechnology, Inc); CD45-Alexa Fluor and CD29-Alexa Fluor (BioLegend); HLA Class 1 ABC (Abcam); and CD34-APC, CD105-APC, CD90-PE, CD73-PE (BD Biosciences).

Techniques: Activation Assay, Transfection, Dominant Negative Mutation, Control, Comparison, Derivative Assay