hdac3  (Sangon Biotech)

Journal: bioRxiv

Article Title: Mitochondrial carrier SLC25A34 links clock, diet, and temperature control of interorganellar lipid cycling

doi: 10.64898/2026.05.30.724257

Figure Lengend Snippet: A, schematic of the multi-omic approach to discover Slc25a34 ; comparing the proteome of BAT from (1) cold-exposed mice (top left, 3 weeks cold exposure vs thermoneutrality, FC>2) and (2) Rev-erbα WKO mice (bottom left, Rev-erbα WKO vs Control, FC>2), (3) cistrome of BAT from Control and Rev-erbα WKO mice (top right, REV-ERBα binding sites located within 10 kb of the nearest gene that show higher occupancy in cold conditions than at thermoneutrality), and (4) BAT transcriptome from cold-exposed Rev-erbα/β dBKO mice (bottom right, genes induced by cold in control mice (FC > 2, FDR < 0.05) and genes induced by double Rev-erbα/β knockout in BAT at thermoneutrality (FC > 2, FDR < 0.05). B , Slc25a34 expression across tissues, fold change relative to the expression level in the tissue at 29°C. C , Slc25a34 and Slc25a35 mRNA and, D , protein levels in BAT during cold adaptation, significance indicates effects of cold exposure compared to 29°C. E , Slc25a34 mRNA levels in scWAT during cold adaptation, significance indicates effects of cold exposure compared to 29°C. F , Circadian expression of Slc25a34 and Slc25a35 mRNA levels in control and Rev-erbα WKO mice acclimated to 29°C. G , SLC25A34 and SLC25A35 protein in BAT of control and Rev-erbα WKO mice at 22°C at ZT10. H , Slc25a34 mRNA levels in BAT of control and Rev-erbα WKO mice at 29°C and after 1.5h of 4°C exposure (ZT4 – ZT5.5). I , REV-ERBα occupancy in the proximal Slc25a34 promoter in BAT at 29°C and after 6 h of 4°C exposure of control and Rev-erbα WKO mice (ZT10). J , Slc25a34 mRNA levels in BAT from control and Hdac3 AKO mice at 22°C and after 3h at 4°C. K , Schematic depicting the role of REV-ERBα in coordinating circadian and cold-mediated regulation of Slc25a34 expression. For all panels, data are represented as mean ±SEM, p < 0.05 = *, p < 0.01 = **, p < 0.001 = ***, p < 0.0001 = ****, unpaired two-tailed multiple t-tests (B, H), One-way ANOVA with the Benjamini, Krieger, and Yekutieli two-stage linear step-up procedure for multiple comparisons correction (C-E), Two-way ANOVA with the Benjamini, Krieger, and Yekutieli two-stage linear step-up procedure for multiple comparisons correction (F, G, J).

Article Snippet: To generate the mouse strain with the adipose-specific Hdac3 knockout ( Hdac3 AKO), Hdac3 f/f mice maintained on a C57BL/6 background were bred to B6;FVB-Tg(Adipoq-Cre)1Evdr/J mice form Jackson laboratory (stock No: 010803) ( ).

Techniques: Control, Binding Assay, Knock-Out, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Mitochondrial carrier SLC25A34 links clock, diet, and temperature control of interorganellar lipid cycling

doi: 10.64898/2026.05.30.724257

Figure Lengend Snippet: A , Slc25a35 expression across tissues, fold change relative to the expression level in the tissue at TN. B , Slc25a35 mRNA levels in scWAT during cold adaptation. C , Slc25a35 mRNA levels in BAT of control and Rev-erbα KO mice at TN and cold. D , Slc25a35 mRNA levels in BAT from control and Hdac3 AKO mice at TN and after 6 h of cold exposure (ZT10). For all panels, error bars represent ±SEM, p < 0.05 = *, p < 0.01 = **, p < 0.001 = ***, p < 0.0001 = ****, unpaired two-tailed multiple t-tests (A), One-way ANOVA (B) and Two-way ANOVA (C, D) with the Benjamini, Krieger, and Yekutieli two-stage linear step-up procedure for multiple comparisons correction.

Article Snippet: To generate the mouse strain with the adipose-specific Hdac3 knockout ( Hdac3 AKO), Hdac3 f/f mice maintained on a C57BL/6 background were bred to B6;FVB-Tg(Adipoq-Cre)1Evdr/J mice form Jackson laboratory (stock No: 010803) ( ).

Techniques: Expressing, Control, Two Tailed Test

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