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: 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