tg2 in 1  (MedChemExpress)

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: Experimental workflow for imaging mass cytometry of fixed spinal cord tissue. Single-cell, spatially resolved, highly multiplexed comparative analysis of the injured spinal cord tissue in TG2 intact and knockout mice was conducted using cytometry by time-of-flight (CyTOF) to evaluate changes in protein expression after SCI. High-dimensional glial and neural phenotypes were visualized using t-distributed stochastic neighbor embedding (t-SNE), a dimensionality reduction technique, to compare target protein expression and cell-type abundance between TG2 intact and knockout tissues following SCI. These approaches enabled unbiased identification of distinct neuronal and glial subpopulations and expression of TG2-specific interactome in modulating cellular responses to SCI. Figure created using BioRender ( https://biorender.com )

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Imaging, Mass Cytometry, Knock-Out, Cytometry, Expressing

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 expression in different neural cell types of the spinal cord under physiological (naïve) and SCI conditions. Representative confocal immunofluorescence images were taken from transverse spinal cord sections of naïve (uninjured) controls (top rows), 24 h post-SCI (middle rows), and 1-week post-SCI (bottom rows). Nuclei were counterstained with Hoechst (blue). A – F Astrocytes labeled with GFAP (green) and TG2 (red in panels B , D , F ). G – L Oligodendrocytes labeled with CC1 (green) and TG2 (red in panels H , J , L ). M – R Neurons labeled with NeuN (green) and TG2 (red in panels N , P , R ). S – X Microglia labeled with Iba1 (green) and TG2 (red in panels T , V , X ). At 24 h and 1-week post-injury, TG2 immunoreactivity increases in all major cell types of the injured spinal cord relative to naïve controls, though is more robust in astrocytes and neurons, reflecting a dynamic regulation after SCI. Scale bars = 50 µm. Quantification ( Y1 – Y4 ): Bar graphs depict mean ± SEM TG2 immunoreactivity (× 10 5 arbitrary units (a.u.)) in GFAP + astrocytes ( Y1 ), CC1 + oligodendrocytes ( Y2 ), NeuN + neurons ( Y3 ), and Iba1 + microglia ( Y4 ) in naïve, at 1-day, and 7-days post-SCI (n = 3 animals per time point; Z depicts the thoracic spinal cord region from which transverse sections were obtained for representative imaging in naïve controls and in the peri-lesional area caudal to the injury at 24 h and 1 week post-SCI). ** p < 0.001, *** p < 0.0001; ns = not significant

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Expressing, Immunofluorescence, Labeling, Imaging

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 knockout enhances functional recovery and tissue preservation following SCI. Behavioral and histological assessments were performed to determine the effects of TG2 KO compared to TG2 intact mice following SCI. A Basso Mouse Scale (BMS) open-field locomotion scores, measured weekly up to 8 weeks post-SCI, demonstrated significantly improved functional recovery in TG2 KO mice compared to TG2 intact mice. B Quantification of errors on the horizontal ladder footfall test revealed a trend toward improved motor coordination in TG2 KO mice at 1- and 2-months post-injury (MPI) compared to injured TG2 intact mice. C – E Histological analysis conducted along the spinal cord axis at 2 months post-SCI revealed: C Significantly reduced lesion area in TG2 KO mice compared to TG2 intact controls. D , E Increased preservation of white matter ( D ) and gray matter ( E ) areas in TG2 KO spinal cords relative to TG2 intact mice, particularly around the injury epicenter and adjacent regions. F – G Representative histological images of spinal cord sections (Luxol fast blue and hematoxylin staining) illustrate greater preservation of tissue integrity in TG2 KO mice ( G ) compared to TG2 intact mice ( F ) at rostral (1.8 mm), injury epicenter, and caudal (1.8 mm) regions. Data are presented as mean ± SEM; statistical significance is indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, *** p < 0.0001, ns = not significant. These combined results demonstrate that TG2 KO significantly promotes functional recovery and histological preservation after SCI

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Knock-Out, Functional Assay, Preserving, Staining

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 knockout reduces fibronectin and chondroitin sulfate proteoglycan deposition in the injured spinal cord. Representative immunofluorescent micrographs from the lesion site at 8 weeks post‐SCI in TG2 intact (top two rows) versus TG2 KO mice (bottom two rows). A , B TG2 intact mouse spinal cord stained for fibronectin (FN, red) and the microglial marker Iba1 (green). C , D TG2 KO spinal cord stained for FN (red) and Iba1 (green). E , F TG2 intact mouse spinal cord stained for the CSPG marker CS56 (green) and GFAP (red). G , H TG2 KO spinal cord similarly stained for CS56 (green) and GFAP (red). Panels ( I ) and ( J ) show quantified immunoreactive (IR) signal intensities (arbitrary units) for FN and CS56, respectively, in control (blue bars) and TG2 KO (orange bars) mice. Results are presented as mean ± SEM. * p < 0.05, ** p < 0.01. Scale bar = 100 µm

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Knock-Out, Staining, Marker, Control

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO enhances axon growth intralesionally after SCI. Representative confocal micrographs of the lesion region at 8 weeks post‐SCI in TG2 intact (top row; A – D ) and TG2 KO (bottom row; E – H ) mice. Sections were immunostained for neurofilament‐H (NF-H; yellow; A , E ), MAP2 (blue; B , F ), GAP-43 (green; C , G ), and serotonin (5HT; red; D , H ). Quantification of immunoreactive (IR) signal intensities is shown in panels I–L : ( I ) NF-H, ( J ) MAP2, ( K ) GAP-43, and ( L ) 5HT. Data represents mean ± SEM. * p < 0.05, ** p < 0.01 vs. control, ns = not significant. Scale bar = 50 µm

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Control

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: IMC analysis reveals TG2 expression and cellular distribution after SCI. Representative IMC images from spinal cord sections at 2 months post-injury are shown for TG2 intact ( B , C ) and KO mice ( D , E ). Samples were acquired using the MCD Viewer (Fluidigm). Panel ( A ) shows the region of the spinal cord section (4800 µm caudal to the SCI center) examined by CyTOF. Panels ( B and D ) illustrate the presence and absence of TG2 expression (red) in spinal cord sections of TG2 intact and KO mice, respectively after SCI, with a clearly delineated gray matter region outlined by dashed white lines. Panels ( C and E ) show multiplexed IMC staining, highlighting various cell markers: neurons (NeuN, cyan), oligodendrocytes (APC, blue), astrocytes (GFAP, green), microglia/macrophages (Iba1, yellow), and endothelial cells (CD31, red). Panel ( F ) quantifies cell-specific TG2 expression across different cell populations. Results indicate robust and relatively uniform TG2 expression in microglia/macrophages (Iba1), astrocytes (GFAP), oligodendrocytes (APC), neurons (NeuN), and endothelial cells (CD31). Data are expressed as mean ± SEM. Scale bar represents 100 µm. Panels G – Z present high-resolution, cell-type-specific views of TG2 distribution in TG2 intact and TG2 KO spinal cord sections, acquired by IMC and visualized in the Fluidigm MCD Viewer. For each marker, the panel in the left image shows the full transverse section, and the right image is a digital zoom of the boxed region from the left panel, highlighting co-localization of TG2 (red) with each of the cell markers (blue): Neurons ( G – J ): NeuN (blue) labels neuronal nuclei. In panels G and H, TG2 (red) is abundant throughout the gray matter neuronal cytoplasm in TG2-intact mice; in panels I and J, the absence of TG2 signal confirms specificity of the antibody in TG2 KO tissue. Astrocytes ( K–N ): GFAP (blue) marks astrocyte processes. TG2 (red) colocalizes extensively with GFAP + astrocytes in TG2 intact mice ( B , L ), whereas only background signal is seen in TG2 KO sections ( M , N ). Microglia/Macrophages ( O – R ): Iba1 (blue) identifies microglia/macrophages. TG2 (red) overlaps with Iba1 + cell bodies and processes in TG2 intact tissue ( O , P ) but is absent in TG2 KO samples ( Q , R ). Oligodendrocytes ( S – V ): APC (blue) labels mature oligodendrocytes. Co-staining ( S , T ) shows widespread TG2 (red) in APC + cells of TG2 intact cords; no TG2 is detected in the KO counterparts ( U , V ). Similarly, endothelial cells ( W – Z ): CD31 (blue) marks the vascular endothelium. TG2 (red) expression decorates CD31 + vessel profiles in TG2 intact sections ( W , X ) but not in TG2 KO tissue ( Y , Z ). Each pair of full-section and magnified panels uses the same scale. Co-localization of TG2 and each cell marker yields a magenta signal in TG2 intact spinal cords, which is absent in the TG2-KO samples. These images collectively demonstrate that TG2 is robustly and uniformly expressed across all major spinal cord cell populations following SCI, and that this signal is abolished in the knockout model

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Expressing, Staining, Marker, Knock-Out

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO modulates oxidative and ER stress as well as apoptotic signaling pathways after SCI. IMC revealed differences in protein expression related to oxidative stress, ER stress, apoptosis, and pro-survival mediators between TG2 intact and KO injured spinal cord tissues at 2 months post-SCI. Representative IMC images from the injured spinal cord sections at 4.8 mm caudal to the injury highlight alterations in multiple signaling factors were acquired using the MCD Viewer (Fluidigm). Panels A – D : Oxidative and ER stress markers, phosphorylated Nrf2 (pNrf2, red), GRP78 (green), and HSP70 (blue), are shown. Panels A and B represent spinal cord images for TG2 + ( A ) and TG2 KO ( B ) animals, respectively. Panels C and D show higher magnifications of the outlined regions, illustrating reduced expression of stress markers in TG2 KO tissues. Panels E – H : Expression of apoptotic pathway factors BNIP3 (red) and Cytochrome C (CytoC, blue) in TG2 + ( E , G ) and TG2 KO ( F , H ) spinal cord sections. Higher-magnification views ( G , H ) highlight differences in apoptotic signaling marker intensity within injured regions. Panels I–L : Pro-survival signaling markers phosphorylated REL-NFκB (pREL-NFκB, red) and apoptosis mediator phosphorylated ERK1/2 (p-ERK1/2, green) are compared between TG2 + ( I , K ) and TG2 KO ( J , L ) groups. Panels K and L show enhanced detail, illustrating differences in inflammation-related signaling activity. The gray matter regions are delineated by dashed white outlines. Results suggest TG2 KO impacts multiple signaling pathways associated with injury progression, potentially favoring a pro-survival and a neuroprotective environment post-SCI. Panels M and N present a quantitative comparison of target protein intensities in TG2 intact and TG2-KO mice (n = 3 per group) at two months post-SCI. Each box plot summarizes the single-cell distribution of metal-tagged antibody signals across the entire neural cell population in injured spinal cord sections, enabling direct side-by-side assessment of key stress, apoptotic, and survival mediators in TG2 intact ( M ) versus TG2 KO ( N ) tissues. Each dot represents the intensity of a metal-tagged antibody in an individual cell, and the overlaying box plots indicate the median, interquartile range, and whiskers (1.5 × IQR). Proteins analyzed include markers associated with cell stress (GRP78, HSP70), oxidative stress (PRDX1, pNrf2), apoptosis (cleaved caspase-3, Bax, Noxa, P53), survival (Bcl-2), and inflammatory signaling (pREL-NFκB, pERK). Ubiquitin levels were also assessed. While some proteins such as GRP78, HSP70, and PRDX1 showed increased expression in TG2 intact tissue, others, including cleaved caspase-3, Bax, Noxa, and P53 did not show significant differential expression between genotypes at this spinal cord level caudal to the lesion. The X-axis labels ( M , N ) correspond to the metal isotopes used to tag each antibody: Sm(149)_CytoC (Samarium-149 for cytochrome c), Sm(150)_pERK (Samarium-150 for pERK1/2), Eu(151)_p53 (Europium-151 for p53), Sm(152)_Ubiquitin (Samarium-152 for ubiquitin), Gd(156)_Bax (Gadolinium-156 for Bax), Tb(159)_HSP70 (Terbium-159 for HSP70), Dy(162)_BNIP3 (Dysprosium-162 for BNIP3), Ho(165)_PRDX1 (Holmium-165 for PRDX1), Er(170)_GRP78 (Erbium-170 for GRP78/BiP), Yb(172)_Casp3 (Ytterbium-172 for cleaved caspase-3), Yb(174)_ pNrf2 (Ytterbium-174 for pNrf2), Lu(175)_Noxa (Lutetium-175 for Noxa), and Yb(176)_pRelA-NFκB (Ytterbium-176 for pRelA/NFκB p65). Panels M and N highlight how TG2 deletion alters the expression of multiple stress-, apoptosis-, and survival-related proteins at the single-cell level. Error bars indicate standard error of the mean. Each dot represents an individual cell-level measurement. Scale bar = 100 µm

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Protein-Protein interactions, Expressing, Marker, Activity Assay, Comparison, Ubiquitin Proteomics, Quantitative Proteomics

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO exhibited reduced pNrf2 levels in the injured spinal cord compared to TG2 intact controls. Representative IMC images of spinal cord sections at 8 weeks post-injury with pNrf2 (pNrf2, red) in TG2 intact and knockout mice A , B were acquired using the MCD Viewer (Fluidigm). Tissue sections are co-stained for NeuN (neurons, green) and GFAP (astrocytes, blue). TG2 intact sections demonstrate extensive pNrf2, co-localized prominently with NeuN and GFAP, whereas pNrf2 is almost undetectable in TG2 KO sections. t-SNE plot generated from three biological replicates per genotype, illustrating single-cell analysis of pNrf2-positive cell populations (dark teal) ( C , D ). TG2 intact spinal cords (bottom half of tSNE) display dense islands of pNrf2-positive cells after SCI that was present ubiquitously in all cell populations, whereas TG2 KO samples (top half of tSNE) exhibited significantly reduced numbers of pNrf2-positive cells. ( C ) The marked reduction of pNrf2-positive cells in TG2 KO mice compared to TG2 intact mice was consistent across all replicates ( F ). Quantitative analysis of pNrf2 expressions in GFAP astrocytes ( D ) and NeuN neurons ( E ) with dramatic reductions in TG2 KO mice after SCI compared to TG2 intact controls (** p < 0.0001)

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Knock-Out, Staining, Generated, Single-cell Analysis

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO markedly reduces GRP78 expression in both astrocytes and neurons of the injured spinal cord. ( A , B ): Representative IMC images of transverse spinal cord sections from TG2 intact ( A ) and KO mice ( B ) mice labeled for GFAP (green), NeuN (blue), and GRP78 (red) were acquired using the MCD Viewer (Fluidigm). ( C ) tSNE plot of combined replicates demonstrates fewer GRP78-positive cells (teal dots) in TG2 KO mice (top half of tSNE, n = 3) compared to TG2 intact mice (bottom half of tSNE, n = 3). The reduced frequency of GRP78-positive cells in TG2 KO mice compared TG2 intact mice was consistent amongst replicates. ( F) Quantification of GRP78 expression associated with GFAP astrocytes ( D ) and NeuN neurons ( E ), shows a significant reduction in the KO group (** p < 0.0001). Data are expressed as mean ± SEM

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Expressing, Labeling

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO reduces HSP70 expression in the injured spinal cord. A , B Representative IMC images of transverse spinal cord sections from TG2 intact ( A ) and TG2 KO mice ( B ) stained for NeuN (green), HSP70 (red), and GFAP (blue) were acquired using the MCD Viewer (Fluidigm). HSP70 IR after SCI was markedly reduced in TG2 KO spinal cords compared to TG2 intact controls ( C ). tSNE analysis showing HSP70-positive cells (teal dots) in TG2 intact (bottom half of tSNE, n = 3) and KO (top half of tSNE n = 3) spinal cords, illustrating a widespread reduction in HSP70 expression in the absence of TG2. D Quantification of HSP70 expression revealed a significant decrease in TG2 KO spinal cords compared to TG2 intact controls (**** p < 0.0001). Data are presented as mean ± SEM

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Expressing, Staining

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO led to reduced cytochrome c expression in the injured spinal cord. ( A , B ) Representative transverse spinal cord sections from TG2 intact ( A ) and KO mice ( B ) were immunostained for NeuN (green), cytochrome c (CytoC, red), and GFAP (blue) were acquired using the MCD Viewer (Fluidigm). ( C , D ) Higher-magnification views of the boxed regions in panels A and B respectively, revealing robust neuronal CytoC staining in TG2 intact spinal cords after SCI but markedly reduced CytoC in KO sections, with the reductions most evident within ventral horn motor neurons. ( E ) tSNE plot of combined data demonstrates similar numbers of CytoC-positive cells in TG2 KO and TG2 intact spinal cords (n=3 per genotype). ( F ) Quantification of CytoC expression within all cells indicates a significant decrease in TG2 KO spinal cords after SCI (****p < 0.0001). Data are expressed as mean ± SEM. Scale bars, 100 μm ( A , B ) and 20 μm ( C , D )

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Expressing, Staining

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: Mass cytometry revealed enhanced phosphorylation of REL-NFκB (pREL-NFκB) in spinal cord tissue after SCI and TG2 KO. A , B Representative immunofluorescence images of spinal cord sections from TG2 intact and KO SCI mice, stained for NeuN (green), pREL-NFκB (red), and GFAP (blue) were acquired using the MCD Viewer (Fluidigm). Scale bars-100 µm. C , D Magnified insets from panels A and B highlight differences in pREL-NFκB immunoreactivity within NeuN neurons in TG2 intact (panel C) versus TG2 KO (panel D) tissues. Scale bars, 20 µm. E tSNE analysis of spinal cord tissue triplicates from TG2 intact and KO mice revealed an increased population of pREL-NFκB-positive cells (teal) in TG2 KO tissues compared to TG2 intact controls. F Quantitative analysis of relative pREL-NFκB IR analyzed globally, indicated significant increases in TG2 KO tissues compared to TG2 intact controls (**** p < 0.0001). Data are presented as mean ± SEM

Article Snippet: Adult, age matched (4–6 months old) TG2 KO and TG2 intact (TG2 +) mice (n = 10–13 per genotype, inclusive of attrition, Total n = 23) or wildtype C57BL6 mice (n = 6–8 per cohort, total n = 14) with a mix of animals from both sexes (The Jackson Laboratory) were used in this study to test the effects of TG2 inhibitors.

Techniques: Mass Cytometry, Phospho-proteomics, Immunofluorescence, Staining

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: Experimental workflow for imaging mass cytometry of fixed spinal cord tissue. Single-cell, spatially resolved, highly multiplexed comparative analysis of the injured spinal cord tissue in TG2 intact and knockout mice was conducted using cytometry by time-of-flight (CyTOF) to evaluate changes in protein expression after SCI. High-dimensional glial and neural phenotypes were visualized using t-distributed stochastic neighbor embedding (t-SNE), a dimensionality reduction technique, to compare target protein expression and cell-type abundance between TG2 intact and knockout tissues following SCI. These approaches enabled unbiased identification of distinct neuronal and glial subpopulations and expression of TG2-specific interactome in modulating cellular responses to SCI. Figure created using BioRender ( https://biorender.com )

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Imaging, Mass Cytometry, Knock-Out, Cytometry, Expressing

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 expression in different neural cell types of the spinal cord under physiological (naïve) and SCI conditions. Representative confocal immunofluorescence images were taken from transverse spinal cord sections of naïve (uninjured) controls (top rows), 24 h post-SCI (middle rows), and 1-week post-SCI (bottom rows). Nuclei were counterstained with Hoechst (blue). A – F Astrocytes labeled with GFAP (green) and TG2 (red in panels B , D , F ). G – L Oligodendrocytes labeled with CC1 (green) and TG2 (red in panels H , J , L ). M – R Neurons labeled with NeuN (green) and TG2 (red in panels N , P , R ). S – X Microglia labeled with Iba1 (green) and TG2 (red in panels T , V , X ). At 24 h and 1-week post-injury, TG2 immunoreactivity increases in all major cell types of the injured spinal cord relative to naïve controls, though is more robust in astrocytes and neurons, reflecting a dynamic regulation after SCI. Scale bars = 50 µm. Quantification ( Y1 – Y4 ): Bar graphs depict mean ± SEM TG2 immunoreactivity (× 10 5 arbitrary units (a.u.)) in GFAP + astrocytes ( Y1 ), CC1 + oligodendrocytes ( Y2 ), NeuN + neurons ( Y3 ), and Iba1 + microglia ( Y4 ) in naïve, at 1-day, and 7-days post-SCI (n = 3 animals per time point; Z depicts the thoracic spinal cord region from which transverse sections were obtained for representative imaging in naïve controls and in the peri-lesional area caudal to the injury at 24 h and 1 week post-SCI). ** p < 0.001, *** p < 0.0001; ns = not significant

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Expressing, Immunofluorescence, Labeling, Imaging

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 knockout enhances functional recovery and tissue preservation following SCI. Behavioral and histological assessments were performed to determine the effects of TG2 KO compared to TG2 intact mice following SCI. A Basso Mouse Scale (BMS) open-field locomotion scores, measured weekly up to 8 weeks post-SCI, demonstrated significantly improved functional recovery in TG2 KO mice compared to TG2 intact mice. B Quantification of errors on the horizontal ladder footfall test revealed a trend toward improved motor coordination in TG2 KO mice at 1- and 2-months post-injury (MPI) compared to injured TG2 intact mice. C – E Histological analysis conducted along the spinal cord axis at 2 months post-SCI revealed: C Significantly reduced lesion area in TG2 KO mice compared to TG2 intact controls. D , E Increased preservation of white matter ( D ) and gray matter ( E ) areas in TG2 KO spinal cords relative to TG2 intact mice, particularly around the injury epicenter and adjacent regions. F – G Representative histological images of spinal cord sections (Luxol fast blue and hematoxylin staining) illustrate greater preservation of tissue integrity in TG2 KO mice ( G ) compared to TG2 intact mice ( F ) at rostral (1.8 mm), injury epicenter, and caudal (1.8 mm) regions. Data are presented as mean ± SEM; statistical significance is indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, *** p < 0.0001, ns = not significant. These combined results demonstrate that TG2 KO significantly promotes functional recovery and histological preservation after SCI

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Knock-Out, Functional Assay, Preserving, Staining

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 knockout reduces fibronectin and chondroitin sulfate proteoglycan deposition in the injured spinal cord. Representative immunofluorescent micrographs from the lesion site at 8 weeks post‐SCI in TG2 intact (top two rows) versus TG2 KO mice (bottom two rows). A , B TG2 intact mouse spinal cord stained for fibronectin (FN, red) and the microglial marker Iba1 (green). C , D TG2 KO spinal cord stained for FN (red) and Iba1 (green). E , F TG2 intact mouse spinal cord stained for the CSPG marker CS56 (green) and GFAP (red). G , H TG2 KO spinal cord similarly stained for CS56 (green) and GFAP (red). Panels ( I ) and ( J ) show quantified immunoreactive (IR) signal intensities (arbitrary units) for FN and CS56, respectively, in control (blue bars) and TG2 KO (orange bars) mice. Results are presented as mean ± SEM. * p < 0.05, ** p < 0.01. Scale bar = 100 µm

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Knock-Out, Staining, Marker, Control

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO enhances axon growth intralesionally after SCI. Representative confocal micrographs of the lesion region at 8 weeks post‐SCI in TG2 intact (top row; A – D ) and TG2 KO (bottom row; E – H ) mice. Sections were immunostained for neurofilament‐H (NF-H; yellow; A , E ), MAP2 (blue; B , F ), GAP-43 (green; C , G ), and serotonin (5HT; red; D , H ). Quantification of immunoreactive (IR) signal intensities is shown in panels I–L : ( I ) NF-H, ( J ) MAP2, ( K ) GAP-43, and ( L ) 5HT. Data represents mean ± SEM. * p < 0.05, ** p < 0.01 vs. control, ns = not significant. Scale bar = 50 µm

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Control

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: IMC analysis reveals TG2 expression and cellular distribution after SCI. Representative IMC images from spinal cord sections at 2 months post-injury are shown for TG2 intact ( B , C ) and KO mice ( D , E ). Samples were acquired using the MCD Viewer (Fluidigm). Panel ( A ) shows the region of the spinal cord section (4800 µm caudal to the SCI center) examined by CyTOF. Panels ( B and D ) illustrate the presence and absence of TG2 expression (red) in spinal cord sections of TG2 intact and KO mice, respectively after SCI, with a clearly delineated gray matter region outlined by dashed white lines. Panels ( C and E ) show multiplexed IMC staining, highlighting various cell markers: neurons (NeuN, cyan), oligodendrocytes (APC, blue), astrocytes (GFAP, green), microglia/macrophages (Iba1, yellow), and endothelial cells (CD31, red). Panel ( F ) quantifies cell-specific TG2 expression across different cell populations. Results indicate robust and relatively uniform TG2 expression in microglia/macrophages (Iba1), astrocytes (GFAP), oligodendrocytes (APC), neurons (NeuN), and endothelial cells (CD31). Data are expressed as mean ± SEM. Scale bar represents 100 µm. Panels G – Z present high-resolution, cell-type-specific views of TG2 distribution in TG2 intact and TG2 KO spinal cord sections, acquired by IMC and visualized in the Fluidigm MCD Viewer. For each marker, the panel in the left image shows the full transverse section, and the right image is a digital zoom of the boxed region from the left panel, highlighting co-localization of TG2 (red) with each of the cell markers (blue): Neurons ( G – J ): NeuN (blue) labels neuronal nuclei. In panels G and H, TG2 (red) is abundant throughout the gray matter neuronal cytoplasm in TG2-intact mice; in panels I and J, the absence of TG2 signal confirms specificity of the antibody in TG2 KO tissue. Astrocytes ( K–N ): GFAP (blue) marks astrocyte processes. TG2 (red) colocalizes extensively with GFAP + astrocytes in TG2 intact mice ( B , L ), whereas only background signal is seen in TG2 KO sections ( M , N ). Microglia/Macrophages ( O – R ): Iba1 (blue) identifies microglia/macrophages. TG2 (red) overlaps with Iba1 + cell bodies and processes in TG2 intact tissue ( O , P ) but is absent in TG2 KO samples ( Q , R ). Oligodendrocytes ( S – V ): APC (blue) labels mature oligodendrocytes. Co-staining ( S , T ) shows widespread TG2 (red) in APC + cells of TG2 intact cords; no TG2 is detected in the KO counterparts ( U , V ). Similarly, endothelial cells ( W – Z ): CD31 (blue) marks the vascular endothelium. TG2 (red) expression decorates CD31 + vessel profiles in TG2 intact sections ( W , X ) but not in TG2 KO tissue ( Y , Z ). Each pair of full-section and magnified panels uses the same scale. Co-localization of TG2 and each cell marker yields a magenta signal in TG2 intact spinal cords, which is absent in the TG2-KO samples. These images collectively demonstrate that TG2 is robustly and uniformly expressed across all major spinal cord cell populations following SCI, and that this signal is abolished in the knockout model

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Expressing, Staining, Marker, Knock-Out

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO modulates oxidative and ER stress as well as apoptotic signaling pathways after SCI. IMC revealed differences in protein expression related to oxidative stress, ER stress, apoptosis, and pro-survival mediators between TG2 intact and KO injured spinal cord tissues at 2 months post-SCI. Representative IMC images from the injured spinal cord sections at 4.8 mm caudal to the injury highlight alterations in multiple signaling factors were acquired using the MCD Viewer (Fluidigm). Panels A – D : Oxidative and ER stress markers, phosphorylated Nrf2 (pNrf2, red), GRP78 (green), and HSP70 (blue), are shown. Panels A and B represent spinal cord images for TG2 + ( A ) and TG2 KO ( B ) animals, respectively. Panels C and D show higher magnifications of the outlined regions, illustrating reduced expression of stress markers in TG2 KO tissues. Panels E – H : Expression of apoptotic pathway factors BNIP3 (red) and Cytochrome C (CytoC, blue) in TG2 + ( E , G ) and TG2 KO ( F , H ) spinal cord sections. Higher-magnification views ( G , H ) highlight differences in apoptotic signaling marker intensity within injured regions. Panels I–L : Pro-survival signaling markers phosphorylated REL-NFκB (pREL-NFκB, red) and apoptosis mediator phosphorylated ERK1/2 (p-ERK1/2, green) are compared between TG2 + ( I , K ) and TG2 KO ( J , L ) groups. Panels K and L show enhanced detail, illustrating differences in inflammation-related signaling activity. The gray matter regions are delineated by dashed white outlines. Results suggest TG2 KO impacts multiple signaling pathways associated with injury progression, potentially favoring a pro-survival and a neuroprotective environment post-SCI. Panels M and N present a quantitative comparison of target protein intensities in TG2 intact and TG2-KO mice (n = 3 per group) at two months post-SCI. Each box plot summarizes the single-cell distribution of metal-tagged antibody signals across the entire neural cell population in injured spinal cord sections, enabling direct side-by-side assessment of key stress, apoptotic, and survival mediators in TG2 intact ( M ) versus TG2 KO ( N ) tissues. Each dot represents the intensity of a metal-tagged antibody in an individual cell, and the overlaying box plots indicate the median, interquartile range, and whiskers (1.5 × IQR). Proteins analyzed include markers associated with cell stress (GRP78, HSP70), oxidative stress (PRDX1, pNrf2), apoptosis (cleaved caspase-3, Bax, Noxa, P53), survival (Bcl-2), and inflammatory signaling (pREL-NFκB, pERK). Ubiquitin levels were also assessed. While some proteins such as GRP78, HSP70, and PRDX1 showed increased expression in TG2 intact tissue, others, including cleaved caspase-3, Bax, Noxa, and P53 did not show significant differential expression between genotypes at this spinal cord level caudal to the lesion. The X-axis labels ( M , N ) correspond to the metal isotopes used to tag each antibody: Sm(149)_CytoC (Samarium-149 for cytochrome c), Sm(150)_pERK (Samarium-150 for pERK1/2), Eu(151)_p53 (Europium-151 for p53), Sm(152)_Ubiquitin (Samarium-152 for ubiquitin), Gd(156)_Bax (Gadolinium-156 for Bax), Tb(159)_HSP70 (Terbium-159 for HSP70), Dy(162)_BNIP3 (Dysprosium-162 for BNIP3), Ho(165)_PRDX1 (Holmium-165 for PRDX1), Er(170)_GRP78 (Erbium-170 for GRP78/BiP), Yb(172)_Casp3 (Ytterbium-172 for cleaved caspase-3), Yb(174)_ pNrf2 (Ytterbium-174 for pNrf2), Lu(175)_Noxa (Lutetium-175 for Noxa), and Yb(176)_pRelA-NFκB (Ytterbium-176 for pRelA/NFκB p65). Panels M and N highlight how TG2 deletion alters the expression of multiple stress-, apoptosis-, and survival-related proteins at the single-cell level. Error bars indicate standard error of the mean. Each dot represents an individual cell-level measurement. Scale bar = 100 µm

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Protein-Protein interactions, Expressing, Marker, Activity Assay, Comparison, Ubiquitin Proteomics, Quantitative Proteomics

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO exhibited reduced pNrf2 levels in the injured spinal cord compared to TG2 intact controls. Representative IMC images of spinal cord sections at 8 weeks post-injury with pNrf2 (pNrf2, red) in TG2 intact and knockout mice A , B were acquired using the MCD Viewer (Fluidigm). Tissue sections are co-stained for NeuN (neurons, green) and GFAP (astrocytes, blue). TG2 intact sections demonstrate extensive pNrf2, co-localized prominently with NeuN and GFAP, whereas pNrf2 is almost undetectable in TG2 KO sections. t-SNE plot generated from three biological replicates per genotype, illustrating single-cell analysis of pNrf2-positive cell populations (dark teal) ( C , D ). TG2 intact spinal cords (bottom half of tSNE) display dense islands of pNrf2-positive cells after SCI that was present ubiquitously in all cell populations, whereas TG2 KO samples (top half of tSNE) exhibited significantly reduced numbers of pNrf2-positive cells. ( C ) The marked reduction of pNrf2-positive cells in TG2 KO mice compared to TG2 intact mice was consistent across all replicates ( F ). Quantitative analysis of pNrf2 expressions in GFAP astrocytes ( D ) and NeuN neurons ( E ) with dramatic reductions in TG2 KO mice after SCI compared to TG2 intact controls (** p < 0.0001)

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Knock-Out, Staining, Generated, Single-cell Analysis

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO markedly reduces GRP78 expression in both astrocytes and neurons of the injured spinal cord. ( A , B ): Representative IMC images of transverse spinal cord sections from TG2 intact ( A ) and KO mice ( B ) mice labeled for GFAP (green), NeuN (blue), and GRP78 (red) were acquired using the MCD Viewer (Fluidigm). ( C ) tSNE plot of combined replicates demonstrates fewer GRP78-positive cells (teal dots) in TG2 KO mice (top half of tSNE, n = 3) compared to TG2 intact mice (bottom half of tSNE, n = 3). The reduced frequency of GRP78-positive cells in TG2 KO mice compared TG2 intact mice was consistent amongst replicates. ( F) Quantification of GRP78 expression associated with GFAP astrocytes ( D ) and NeuN neurons ( E ), shows a significant reduction in the KO group (** p < 0.0001). Data are expressed as mean ± SEM

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Expressing, Labeling

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO reduces HSP70 expression in the injured spinal cord. A , B Representative IMC images of transverse spinal cord sections from TG2 intact ( A ) and TG2 KO mice ( B ) stained for NeuN (green), HSP70 (red), and GFAP (blue) were acquired using the MCD Viewer (Fluidigm). HSP70 IR after SCI was markedly reduced in TG2 KO spinal cords compared to TG2 intact controls ( C ). tSNE analysis showing HSP70-positive cells (teal dots) in TG2 intact (bottom half of tSNE, n = 3) and KO (top half of tSNE n = 3) spinal cords, illustrating a widespread reduction in HSP70 expression in the absence of TG2. D Quantification of HSP70 expression revealed a significant decrease in TG2 KO spinal cords compared to TG2 intact controls (**** p < 0.0001). Data are presented as mean ± SEM

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Expressing, Staining

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: TG2 KO led to reduced cytochrome c expression in the injured spinal cord. ( A , B ) Representative transverse spinal cord sections from TG2 intact ( A ) and KO mice ( B ) were immunostained for NeuN (green), cytochrome c (CytoC, red), and GFAP (blue) were acquired using the MCD Viewer (Fluidigm). ( C , D ) Higher-magnification views of the boxed regions in panels A and B respectively, revealing robust neuronal CytoC staining in TG2 intact spinal cords after SCI but markedly reduced CytoC in KO sections, with the reductions most evident within ventral horn motor neurons. ( E ) tSNE plot of combined data demonstrates similar numbers of CytoC-positive cells in TG2 KO and TG2 intact spinal cords (n=3 per genotype). ( F ) Quantification of CytoC expression within all cells indicates a significant decrease in TG2 KO spinal cords after SCI (****p < 0.0001). Data are expressed as mean ± SEM. Scale bars, 100 μm ( A , B ) and 20 μm ( C , D )

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Expressing, Staining

Journal: Acta Neuropathologica Communications

Article Title: Removal or inhibition of transglutaminase 2 decreases cellular stress, supporting tissue preservation, and recovery after SCI

doi: 10.1186/s40478-025-02135-4

Figure Lengend Snippet: Mass cytometry revealed enhanced phosphorylation of REL-NFκB (pREL-NFκB) in spinal cord tissue after SCI and TG2 KO. A , B Representative immunofluorescence images of spinal cord sections from TG2 intact and KO SCI mice, stained for NeuN (green), pREL-NFκB (red), and GFAP (blue) were acquired using the MCD Viewer (Fluidigm). Scale bars-100 µm. C , D Magnified insets from panels A and B highlight differences in pREL-NFκB immunoreactivity within NeuN neurons in TG2 intact (panel C) versus TG2 KO (panel D) tissues. Scale bars, 20 µm. E tSNE analysis of spinal cord tissue triplicates from TG2 intact and KO mice revealed an increased population of pREL-NFκB-positive cells (teal) in TG2 KO tissues compared to TG2 intact controls. F Quantitative analysis of relative pREL-NFκB IR analyzed globally, indicated significant increases in TG2 KO tissues compared to TG2 intact controls (**** p < 0.0001). Data are presented as mean ± SEM

Article Snippet: Further, TG2 is a multifunctional enzyme, possessing also GTPase, isopeptidase, and protein disulfide isomerase activities, underscoring a diverse set of functions in cell signaling [ , ].

Techniques: Mass Cytometry, Phospho-proteomics, Immunofluorescence, Staining