tgm1  (Proteintech)

Journal: The Journal of Cell Biology

Article Title: Spectrin coordinates cell shape and signaling essential for epidermal differentiation

doi: 10.1083/jcb.202502071

Figure Lengend Snippet: αII-spectrin regulates epidermal differentiation and barrier function. (a) Dorsal skin from shCtr and shSptan1 0595 -transduced E17.5 embryos immunolabeled for the basal layer marker K14, the differentiation marker K10, and the granular layer marker loricrin. Insets show the transduced cells (H2B−GFP+). (b) Quantification of cell area and shape of suprabasal shSptan1 K14 + and K14 - cells in E17.5 embryos. Values of basal and suprabasal shCtr and basal shSptan1 are equal to Lines: Mean values; dots: single cells pooled from 3 embryos with >100 cells for each condition. ****P < 0.0001 with Kruskal–Wallis, Dunn’s multiple comparison test. (c) Immunofluorescence analysis for the TJ marker occludin in Ctr and Sptan1 −/− primary keratinocytes differentiated for 48 h in high Ca 2+ . Representative example of three biological replicates each. (d) Transepithelial resistance (TER) measurements in Ctr and αII-spectrin knockdown keratinocytes after switching to high Ca 2+ . Line represent means over time of three biological replicates each. Representative experiment of n > 10 biological replicates. (e) Newborn epidermal whole-mount immunofluorescence analysis for the TJ marker ZO-1, revealing impaired alignment of the upper old (red arrowheads) and the lower new TJ rings (blue arrowheads) in the granular layer 2 (SG2). Maximum projection of the granular layer. (f) Illustration of cell shapes and TJ organization in the SG2 of Ctr and Sptan1 epi−/− epidermis. (g) Dorsal skin sections from shCtr and shSptan1 0595- transduced E17.5 embryos. Sections were processed for transglutaminase 1 (TGM1) activity assay. Upper Insets show the transduced cells (H2B−RFP+). (h) Quantification of TGM1 intensity from data shown in g. Data are the mean ± SD of 30 ROI from n = 3 embryos per condition. Bars: mean normalized intensity; dots individual microscopy fields. *P = 0.0471 by unpaired t- test. (i) Quantification of TGM1 activity cortical enrichment from the data shown in g. Mean ± SD from 60 individual cells from n = 3 embryos per condition. Bars: TGM1 activity cortex/cytoplasm intensity ratio mean; dots: individual cells. ***P = 0.0003 with Kolmogorov–Smirnov. (j) Dye exclusion assay: shCtr and shSptan1 0595 -transduced E17.5 embryos were treated with toluidine blue dye to evaluate the skin barrier. (k) Dye exclusion assay: Ctr and Sptan1 epi−/− E17.5 embryos were treated with toluidine blue dye to evaluate the skin barrier. (l) Dorsal skin section from Ctr and E-cadherin epi−/− newborn mice. Sections were processed for TGM1 activity assay or negative Ctr (mutated TGM substrate, pepQNK5). (m) Quantification of TGM1 intensity from data shown in l. (n) Transepidermal water loss (TEWL) measurements on Ctr and Sptan1 epi −/− newborn mice. Dots represent individual mice. Data are the mean of 27 fields of view from n = 3 newborn mice per condition. Bars: Mean intensity; dots individual microscopy fields. ****P < 0.0001 with Kolmogorov–Smirnov. ROI, region of interest.

Article Snippet: Ankyrin3 (IF 1:500, WB 1:1,000, #27766-1-AP; Proteintech), E-cadherin (IF 1:200, #610182; BD Transduction Laboratories, clone number 36 or IF 1:500, #3195; Cell Signaling), EGFR (IF 1:500, #ab52894; Abcam), pEGFR (Y1068) (IF 1:500, #ab40815; Abcam), GAPDH (WB 1:10,000, #AM4300; Ambion or WB 1:1,000, #5174; Cell Signaling), GFP (1:3,000, #ab13970; Abcam), Keratin10 (IF 1:1,000, #PRB-159P; BioLegend), Keratin14 (IF 1:2,000, #PRB 155P; Covance), Loricrin (1:1,000, #Poly19051; BioLegend), Myosin heavy chain IIa (IF 1:500, PRB-440; BioLegend), phospho-Myosin Light Chain 2 (Thr18/Ser19) (IF 1:100, #3674; Cell Signaling), occludin (IF 1:400, #33-1,500; Invitrogen), TRPV3 (IF 1:1,000, #b94582; Abcam), αII-Spectrin (IF 1:500, WB 1:1,000, ab11755; Abcam), TGM1 (IF 1:500, #12912-3-AP; Proteintech), α-catenin (IF 1:2,000, #C2081; Sigma-Aldrich), β-catenin (IF 1:1,000, #ab32572; Abcam), and YAP (IF 1:500, #14074; Cell Signaling).

Techniques: Immunolabeling, Marker, Comparison, Immunofluorescence, Knockdown, Activity Assay, Microscopy, Exclusion Assay

Journal: The Journal of Cell Biology

Article Title: Spectrin coordinates cell shape and signaling essential for epidermal differentiation

doi: 10.1083/jcb.202502071

Figure Lengend Snippet: αII-spectrin regulates epidermal differentiation and barrier function. (a) Dorsal skin sections from shSptan1 0595 -transduced E17.5 embryos immunolabeled for the basal layer marker K14 and the suprabasal marker K10. (b) Dorsal skin sections from shSptan1 9753 -transduced E17.5 embryos immunolabeled for the basal layer marker K14 and the granular layer marker loricrin. Insets show the transduced cells (H2B−GFP+). (c) Dorsal skin sections from shSptan1 0595 -transduced E17.5 embryos immunolabeled for the cleavage furrow marker survivin. Yellow lines show representative axes of division. Graph: Quantification of spindle orientation plotted as a cumulative frequency distribution. NS: P = 0.3485 with Mann–Whitney. (d) Dorsal skin sections from shSptan1 0595 -transduced E17.5 embryos immunolabeled for EdU. (e) Quantification of EdU + basal and suprabasal layer cells from the data shown in d. Bars: mean ± SD from n = 3 embryos per condition. Dots: average EdU + basal and suprabasal layer cells from each embryo with Mann–Whitney. (f) Quantification of cell (nuclei) numbers from primary Ctr and Sptan1 −/− or Sptan1 siRNA-treated keratinocytes differentiated for 48 h in high Ca 2+ . Dots: Mean values from biological replicates. >360 cells counted for Ctr/ Sptan1 −/− each and >20,000 cells for siCtr/ siSptan1 each with Mann–Whitney. (g) Dorsal skin section from Ctr and Sptan1 epi−/− newborn mice. Sections were processed for transglutaminase 1 (TGM1) activity assay or negative Ctr (mutated TGM substrate, pepQNK5). (h) Quantification of TGM1 intensity from data shown in g. Data are the mean of 30 fields of view from n = 3 newborn mice per condition. Bars: Mean intensity; dots individual microscopy fields. ****P < 0.0001 with Kolmogorov–Smirnov. (i) Dorsal skin sections from Ctr, Ecad epi−/− and Sptan1 epi−/− newborn mice immunolabeled for total TGM1 protein. (j and k) Quantification of TGM1 intensity in Ctr, Ecad epi−/− and Sptan1 epi−/− . Lines: Mean values/biological replicate. Nonsignificant with Mann–Whitney. (l) Dye exclusion assay: shCtr and shSptan1 9753 -transduced E17.5 embryos were treated with toluidine blue dye to evaluate the skin barrier. (m) Dye exclusion assay: shCtr and shSptan1 0595 -transduced E18.5 embryos were treated with toluidine blue.

Article Snippet: Ankyrin3 (IF 1:500, WB 1:1,000, #27766-1-AP; Proteintech), E-cadherin (IF 1:200, #610182; BD Transduction Laboratories, clone number 36 or IF 1:500, #3195; Cell Signaling), EGFR (IF 1:500, #ab52894; Abcam), pEGFR (Y1068) (IF 1:500, #ab40815; Abcam), GAPDH (WB 1:10,000, #AM4300; Ambion or WB 1:1,000, #5174; Cell Signaling), GFP (1:3,000, #ab13970; Abcam), Keratin10 (IF 1:1,000, #PRB-159P; BioLegend), Keratin14 (IF 1:2,000, #PRB 155P; Covance), Loricrin (1:1,000, #Poly19051; BioLegend), Myosin heavy chain IIa (IF 1:500, PRB-440; BioLegend), phospho-Myosin Light Chain 2 (Thr18/Ser19) (IF 1:100, #3674; Cell Signaling), occludin (IF 1:400, #33-1,500; Invitrogen), TRPV3 (IF 1:1,000, #b94582; Abcam), αII-Spectrin (IF 1:500, WB 1:1,000, ab11755; Abcam), TGM1 (IF 1:500, #12912-3-AP; Proteintech), α-catenin (IF 1:2,000, #C2081; Sigma-Aldrich), β-catenin (IF 1:1,000, #ab32572; Abcam), and YAP (IF 1:500, #14074; Cell Signaling).

Techniques: Immunolabeling, Marker, MANN-WHITNEY, Activity Assay, Microscopy, Exclusion Assay

Journal: The Journal of Cell Biology

Article Title: Spectrin coordinates cell shape and signaling essential for epidermal differentiation

doi: 10.1083/jcb.202502071

Figure Lengend Snippet: EGFR activity regulates cortical TRPV3 localization. (a) Dorsal skin sections from shCtr; TRPV3-GFP–transduced E17.5 embryos treated with DMSO or gefitinib immunolabeled for αII-spectrin and pEGFR. (b and c) Quantification of TRPV3-GFP and αII-spectrin cortical enrichment from the data shown in a. Mean ± SD from 30 individual cells from n = 3 embryos per condition. Bars: TRPV3-GFP cortex/cytoplasm ratio; dots: individual cells. **P = 0.0029 for TRPV3-GFP and ***P = 0.0001 for αII-spectrin with Kolmogorov-Smirnov. (d and f) Primary mouse keratinocytes cultured in high-calcium (1.5 mM) medium treated with DMSO, gefitinib, or TGF-α and immunolabelled for p-EGFR, TRPV3, and αII-spectrin. Boxes indicate the location of the magnified area. (e and g) Quantification of TRPV3 and αII-spectrin intensity from the data shown in d and f. Mean ± SD from ∼200 mature junctions from n = 3 experiment per condition. Bars: mean normalized intensity; dots: individual junctions. Nuclei were stained with DAPI. **P = 0.001 and ****P > 0.0001 for TRPV3 intensity. ****P > 0.0001 and ***P = 0.0001 for αII-spectrin intensity with Kolmogorov–Smirnov. (h) Dorsal skin sections from E17.5 wild-type embryos treated with DMSO and gefitinib. Sections were processed for transglutaminase 1 (TGM1) activity assay. (i) Quantification of crosslinked TGM substrate intensity. Mean ± SD of 30 ROIs from n = 3 embryos per condition. Bars: mean normalized intensity; dots: individual microscopy fields. NS: P = 0.3876 with Kolmogorov–Smirnov. (j) Quantification of cortical enrichment of crosslinked TGM substrate. Mean ± SD from 60 individual cells from n = 3 embryos per condition. Bars: means of cortex/cytoplasm ratio; dots: individual cells. ****P < 0.0001 with Kolmogorov–Smirnov. Nuclei were stained with DAPI; dashed lines indicate the dermal-epidermal border. (k) Model - High-tension spectrin-actomyosin cortices regulate TGM activity. Illustration of junction and cytoskeleton distribution across epidermal layers, with spectrin most enriched in SG3 and F-actin most enriched in SG1. Lower left: illustration of spectrin and myosin-dependent organization of cortical F-actin. Upper left: Working model of how lattice organization and myosin tension regulate EGFR/TRPV3 signaling complexes, resulting in Ca 2+ influx and cortical TGM activation. ROI, region of interest.

Article Snippet: Ankyrin3 (IF 1:500, WB 1:1,000, #27766-1-AP; Proteintech), E-cadherin (IF 1:200, #610182; BD Transduction Laboratories, clone number 36 or IF 1:500, #3195; Cell Signaling), EGFR (IF 1:500, #ab52894; Abcam), pEGFR (Y1068) (IF 1:500, #ab40815; Abcam), GAPDH (WB 1:10,000, #AM4300; Ambion or WB 1:1,000, #5174; Cell Signaling), GFP (1:3,000, #ab13970; Abcam), Keratin10 (IF 1:1,000, #PRB-159P; BioLegend), Keratin14 (IF 1:2,000, #PRB 155P; Covance), Loricrin (1:1,000, #Poly19051; BioLegend), Myosin heavy chain IIa (IF 1:500, PRB-440; BioLegend), phospho-Myosin Light Chain 2 (Thr18/Ser19) (IF 1:100, #3674; Cell Signaling), occludin (IF 1:400, #33-1,500; Invitrogen), TRPV3 (IF 1:1,000, #b94582; Abcam), αII-Spectrin (IF 1:500, WB 1:1,000, ab11755; Abcam), TGM1 (IF 1:500, #12912-3-AP; Proteintech), α-catenin (IF 1:2,000, #C2081; Sigma-Aldrich), β-catenin (IF 1:1,000, #ab32572; Abcam), and YAP (IF 1:500, #14074; Cell Signaling).

Techniques: Activity Assay, Immunolabeling, Cell Culture, Staining, Microscopy, Activation Assay

Journal: Cell Death & Disease

Article Title: Transglutaminase 2 exacerbates ovarian cancer survival by directly inactivating GSK3β

doi: 10.1038/s41419-026-08447-0

Figure Lengend Snippet: A Quantification of histological (H) scores for transglutaminase 2 (TGase 2) in normal ovarian tissue ( n = 27) and primary ovarian tumors ( n = 180) using inForm™ image‑analysis software. Scale bar, 100 µm. B Quantitative data for TGase 2 staining on tissue microarrays from normal controls ( n = 27) and tumors classified as stage I ( n = 83), stage II ( n = 24), stage III–IV ( n = 37), or metastatic lesions ( n = 36). Representative immunohistochemical (IHC) images are shown Supplementary Fig. . Scale bar, 100 µm. C Correlation analysis between TGM2 and epithelial-to-mesenchymal transition (EMT)–related genes grouped by biological function. The graphs of TMA analysis are presented as mean ± standard error of the mean (SEM). More information on this cohort is provided in Supplementary Fig. . Comparisons between two groups were performed using the Mann–Whitney test, while one-way analysis of variance (ANOVA) was used for comparisons among three or more groups. Statistical significance was defined as * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Fixed cells were incubated overnight at 4 °C with primary antibodies against TGase 2 (Cat. No. 68006-1-Ig; 1:800 dilution, Proteintech) and GSK3β (Cat. No. #12456; 1:500 dilution, Cell Signaling Technology).

Techniques: Software, Staining, Immunohistochemical staining, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: Transglutaminase 2 exacerbates ovarian cancer survival by directly inactivating GSK3β

doi: 10.1038/s41419-026-08447-0

Figure Lengend Snippet: A TGase 2 modulates ovarian cancer cell migration. OVCAR‑5, OVCAR‑8, and SKOV‑3 cells were transfected for 48 h with TGM2 siRNA (20 or 40 nM), after which their migratory capacity was measured. Simultaneously, OVCAR‑3, OVCAR‑4, and SKOV‑3 cells were transfected with TGM2 expression plasmids (2 or 4 µg) for 48 h and tested in the same assay. RFUs indicate relative fluorescence units. B Effect of TGase 2 on epithelial-to-mesenchymal transition (EMT) markers. OVCAR‑8 cells transfected with TGM2 siRNA (40 nM) and SKOV‑3 cells transfected with TGM2 plasmid (4 µg, each for 48 h) were analyzed by immunoblotting for EMT-associated proteins. C WNT/β‑catenin signaling in ovarian cancer cells. OVCAR‑8 and SKOV‑3 cells were treated with lithium chloride (LiCl; 30 mM, 8 h) to inhibit GSK3β and activate the β‑catenin pathway. D TGase 2–GSK3β interaction detected by co‑immunoprecipitation. OVCAR‑8 cells transfected with TGM2 siRNA (40 nM) for 48 h were lysed under serum‑supplemented or serum‑starved conditions before co‑immunoprecipitation. E TGase 2 regulates GSK3β turnover through autophagy. OVCAR‑8 cells transfected with TGM2 siRNA (40 nM) for 48 h were starved and then treated with chloroquine (CQ; 50 µM, 6 h) or left untreated, followed by immunoblot analysis. All graphs are displayed as mean ± standard deviation (SD). All experiments were performed with three samples per group ( n = 3). Comparisons between two groups were performed using the Student’s t test, whereas comparisons among three or more groups were conducted using one-way analysis of variance (ANOVA). Statistical significance was defined as * p < 0.05 or ** p < 0.01 or *** p < 0.001, and ns indicates not significant.

Article Snippet: Fixed cells were incubated overnight at 4 °C with primary antibodies against TGase 2 (Cat. No. 68006-1-Ig; 1:800 dilution, Proteintech) and GSK3β (Cat. No. #12456; 1:500 dilution, Cell Signaling Technology).

Techniques: Migration, Transfection, Expressing, Fluorescence, Plasmid Preparation, Western Blot, Standard Deviation

Journal: Cell Death & Disease

Article Title: Transglutaminase 2 exacerbates ovarian cancer survival by directly inactivating GSK3β

doi: 10.1038/s41419-026-08447-0

Figure Lengend Snippet: A N-terminal region of TGase 2 is required for its interaction with GSK3β. HEK-293 cells were co-transfected with plasmids encoding FLAG-GSK3β and either wild-type TGM2 or the indicated TGM2 deletion mutants (Δ2-139, Δ147-460, Δ472-583, Δ592-673). Complexes were immunoprecipitated using an anti-FLAG antibody and analyzed by immunoblotting. B TGase 2 binds the catalytic core of GSK3β. HEK-293 cells were transfected with plasmids encoding FLAG-TGM2 and either wild-type GSK3β or point-mutated variants (Y216F, V267G/E268R, or F291L). Protein complexes were purified via anti-FLAG immunoprecipitation and examined by immunoblot analysis. C In silico docking predicts a direct interaction between TGase 2 and GSK3β. ClusPro docking of GSK3β (PDB 4NM5, gray) with TGase 2 (PDB 2Q3Z, pink) is shown as cartoons; interface residues are displayed as sticks (GSK3β, gray; TGase 2, blue-green and yellow). D TGase 2 inhibits the kinase activity of GSK3β in a dose-dependent manner. Recombinant GSK3β (2 ng) was incubated with increasing amounts of rhTGase 2 (0–8 ng), and kinase activity was quantified. The graphs display mean ± standard deviation (SD). Assays were performed with three samples per group ( n = 3). rhTGase 2; recombinant human TGase 2. E The TGase 2:GSK3β ratio modulates competing protein-protein interactions. Elevated TGase 2 expression strengthens the TGase 2–GSK3β interaction while reducing the association between GSK3β and β‑catenin. F Domain organization of GSK3β and TGase 2. Schematic diagrams depict the primary structures and domains of each protein, with residue ranges corresponding to the constructs analyzed in this study. BD: substrate binding domain.

Article Snippet: Fixed cells were incubated overnight at 4 °C with primary antibodies against TGase 2 (Cat. No. 68006-1-Ig; 1:800 dilution, Proteintech) and GSK3β (Cat. No. #12456; 1:500 dilution, Cell Signaling Technology).

Techniques: Transfection, Immunoprecipitation, Western Blot, Purification, In Silico, Activity Assay, Recombinant, Incubation, Standard Deviation, Protein-Protein interactions, Expressing, Residue, Construct, Binding Assay

Journal: Cell Death & Disease

Article Title: Transglutaminase 2 exacerbates ovarian cancer survival by directly inactivating GSK3β

doi: 10.1038/s41419-026-08447-0

Figure Lengend Snippet: A Generation of TGM2 knock-out cell lines using the CRISPR system. TGase 2 expression was confirmed by immunoblotting, and clone #2, which showed the lowest TGase 2 level, was selected for in vivo studies. To mimic ovarian cancer growth and metastasis, we evaluated antitumor efficacy using an intraperitoneal orthotopic model. Loss of TGase 2 decreases intraperitoneal (i.p.) tumor growth in an OVCAR‑8/Luc xenograft model. BALB/c‑nu/nu mice ( n = 12 per group) were inoculated i.p. (intraperitoneally) with 1 ×10 7 parental OVCAR‑8/Luc cells or OVCAR‑8/Luc cells with TGM2 KO. Tumor burden was monitored weekly via in vivo bioluminescence imaging (IVIS) and measured as total photon flux using Living Image software. Graphs show mean ± standard deviation (SD). Comparisons between groups used a Two-way ANOVA. Statistical significance was set at *** p < 0.001. ROI, region of interest. B Kaplan–Meier survival curves for the cohorts in ( A ). Deletion of TGM2 significantly improved overall survival (** p < 0.01, log‑rank test). To examine if TGase 2 also plays a role in late metastatic stages, we used a tail-vein lung metastasis model. TGase 2 promotes experimental ovarian cancer metastasis. Parental or TGM2 ‑KO OVCAR‑8/Luc cells were injected into the tail vein of BALB/c‑nude mice ( n = 4 per group). C Metastatic lesions collected at the endpoint were analyzed through immunohistochemistry. Scale bar, 100 μm (yellow) and 2 mm (black). D Quantification of metastatic burden and nodules. * p < 0.05, ** p < 0.01. E , F Immunohistochemical analysis of GSK3β expression in TG2-deficient tumors was performed using the Vectra Polaris™ Automated Quantitative Pathology Imaging System and inForm software (Akoya Biosciences, Waltham, MA, USA). Scale bar, 100 μm. Bar graphs display the number of metastatic nodules and the total tumor area per mouse (mean ± SD; n = 4). ** p < 0.01.

Article Snippet: Fixed cells were incubated overnight at 4 °C with primary antibodies against TGase 2 (Cat. No. 68006-1-Ig; 1:800 dilution, Proteintech) and GSK3β (Cat. No. #12456; 1:500 dilution, Cell Signaling Technology).

Techniques: Knock-Out, CRISPR, Expressing, Western Blot, In Vivo, Imaging, Software, Standard Deviation, Injection, Immunohistochemistry, Immunohistochemical staining

Journal: Cell Death & Disease

Article Title: Transglutaminase 2 exacerbates ovarian cancer survival by directly inactivating GSK3β

doi: 10.1038/s41419-026-08447-0

Figure Lengend Snippet: A Streptonigrin (STN) disrupts the TGase2–GSK3β interaction. HEK-293 cells were co‑transfected with FLAG- TGM2 and GSK3β -HA plasmids, cultured for 48 h, and then treated with the indicated concentrations of STN for 1 h. B STN reduces intracellular co‑localization of TGase 2 and GSK3β. OVCAR‑8 cells were exposed to 500 nM STN for 1 h and analyzed by confocal microscopy. Scale bar, 20 µm. C To assess whether TGase 2 inhibition suppresses metastasis, we utilized a tail-vein lung metastasis model. TGase2 inhibition decreases metastatic burden in a tail‑vein ovarian cancer model. SKOV‑3 cells were injected into the tail vein of BALB/c‑nude mice ( n = 4 per group). Metastatic lesions were collected and analyzed via immunohistochemistry. Scale bar, 100 µm. Bar graphs represent the number of metastatic nodules and total tumor area (mean ± SD; n = 4). Scale bar, 200 µm. D To confirm whether the same effect is observed in an authentic ovarian cancer metastasis model, we revalidated it using an orthotopic model. TGase2 inhibition delays tumor growth in an intraperitoneal xenograft model. OVCAR‑8/Luc cells (1 × 10⁷) were injected intraperitoneally into mice ( n = 9 per group). Beginning two days before cell inoculation, mice received oral PBS or STN (0.01 or 0.1 mg/kg) five days per week. Tumor growth was monitored weekly by bioluminescence imaging (IVIS) and total photon flux was plotted (mean ± SEM). E Kaplan–Meier survival curves for the mice in Fig. over 80 days after cell injection. All graphs are presented as mean ± standard deviation (SD). Multiple comparisons among three or more groups were performed using one-way analysis of variance (ANOVA). Statistical significance was considered at * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Fixed cells were incubated overnight at 4 °C with primary antibodies against TGase 2 (Cat. No. 68006-1-Ig; 1:800 dilution, Proteintech) and GSK3β (Cat. No. #12456; 1:500 dilution, Cell Signaling Technology).

Techniques: Cell Culture, Confocal Microscopy, Inhibition, Injection, Immunohistochemistry, Imaging, Standard Deviation

Journal: Cell Death & Disease

Article Title: Transglutaminase 2 exacerbates ovarian cancer survival by directly inactivating GSK3β

doi: 10.1038/s41419-026-08447-0

Figure Lengend Snippet: A Conventional chemotherapies increase TGase 2 levels and decrease GSK3β levels. OVCAR‑3 or SKOV‑3 cells were treated with cisplatin (DDP, 1 mM) or paclitaxel (PTX, 10 μM) for 24 h, after which TGase 2 and GSK3β protein levels were measured by immunoblotting. B Chemotherapy treatment enhances the migratory ability of ovarian cancer cells. Cell migration was quantified after treatment with DDP or PTX in OVCAR-3 and SKOV-3 cells. The graphs show mean ± standard deviation (SD). Assays used three samples per group ( n = 3). C To assess metastasis suppression and the anti-tumor effects of first-line ovarian cancer therapy combined with a TGase 2 inhibitor, we performed tests using an orthotopic model using OVCAR‑8/Luc cells. Pharmacological inhibition of TGase 2 works synergistically with conventional chemotherapies and prolongs survival in an ovarian cancer mouse model. Mice ( n ≥ 8 per group) received phosphate-buffered saline (PBS), streptonigrin (STN; 0.01 mg/kg), DDP (1 mg/kg), PTX (1 mg/kg), STN + DDP, or STN + PTX. Kaplan–Meier survival curves are shown; median survival was calculated using Kaplan–Meier statistics. D Quantification of histological scores for GSK3β in ovarian cancer patient tissue using inForm™ image analysis software. Tumors were classified as stage I ( n = 83), stage II ( n = 24), stage III–IV ( n = 37), or metastatic lesions ( n = 36). Representative immunohistochemical (IHC) images are included (Supplementary Fig. ) scale bar, 100 µm. E Comparison of TG2 and GSK3β expression patterns during tumor development and metastasis. The TMA analysis graphs are presented as mean ± standard error of the mean (SEM). Multiple comparisons among three or more groups were performed with one-way analysis of variance (ANOVA). Statistical significance was set at ** p < 0.01, *** p < 0.001, and **** p < 0.0001. ns, not significant.

Article Snippet: Fixed cells were incubated overnight at 4 °C with primary antibodies against TGase 2 (Cat. No. 68006-1-Ig; 1:800 dilution, Proteintech) and GSK3β (Cat. No. #12456; 1:500 dilution, Cell Signaling Technology).

Techniques: Western Blot, Migration, Standard Deviation, Inhibition, Saline, Software, Immunohistochemical staining, Comparison, Expressing

Journal: Molecular Medicine Reports

Article Title: TGM1 as a novel signature gene in psoriasis identified by integrative bioinformatics and experimental validation

doi: 10.3892/mmr.2025.13755

Figure Lengend Snippet: Identification of biomarkers of psoriasis based on machine learning algorithms. (A) Optimal penalization coefficient λ in the LASSO model and (B) average deviance values for each model with a given λ. (C) Association between the total number of trees in the RF algorithm and the error rates, and (D) the top 15 signature genes. (E) Error rate of the curve after 5-fold cross-validation of SVM-RFE algorithm to select the signature genes. Error rate predicted by the 29 genes is 0.0252. (F) Accuracy rate predicted by 29 genes was 0.975. (G) Venn diagram of the biomarkers identified using LASSO, SVM-RFE and RF algorithms. RF, random forest; SVM-RFE, support vector machine-recursive feature elimination; LASSO, least absolute shrinkage and selection operator; TGM1, transglutaminase 1, CV, cross validation.

Article Snippet: Sections were incubated with a primary antibody against TGM1 (1: 100 cat. no. 12912-3-AP; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Biomarker Discovery, Plasmid Preparation, Selection

Journal: Molecular Medicine Reports

Article Title: TGM1 as a novel signature gene in psoriasis identified by integrative bioinformatics and experimental validation

doi: 10.3892/mmr.2025.13755

Figure Lengend Snippet: Identification of biomarkers of psoriasis. (A) TGM1 and its connected genes in the protein-protein interaction network constructed using 163 differentially expressed genes with |log 2 (FC)|>3. (B) Box plot of TGM1 expression in LS and NLS tissues of patients with psoriasis. (C) A receiver operating characteristic curve was employed to assess the diagnosis relevance of TGM1 in psoriasis. LS, lesional skin; NLS, non-lesional skin; AUC, area under the curve; TGM1, transglutaminase 1.

Article Snippet: Sections were incubated with a primary antibody against TGM1 (1: 100 cat. no. 12912-3-AP; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Construct, Expressing, Biomarker Discovery

Journal: Molecular Medicine Reports

Article Title: TGM1 as a novel signature gene in psoriasis identified by integrative bioinformatics and experimental validation

doi: 10.3892/mmr.2025.13755

Figure Lengend Snippet: Functional enrichment for TGM1 via single-gene Gene Set Enrichment Analysis. (A) Up- and (B) Downregulated signaling pathways involving TGM1. TGM1, transglutaminase 1.

Article Snippet: Sections were incubated with a primary antibody against TGM1 (1: 100 cat. no. 12912-3-AP; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Functional Assay, Protein-Protein interactions

Journal: Molecular Medicine Reports

Article Title: TGM1 as a novel signature gene in psoriasis identified by integrative bioinformatics and experimental validation

doi: 10.3892/mmr.2025.13755

Figure Lengend Snippet: Validation of TGM1 expression in validation datasets. Volcano plots showing the differentially expressed genes in the (A) GSE53552 and (D) GSE13355 datasets based on |FC|>2 and P<0.05. The dark red dot indicates TGM1. Boxplots showing TGM1 expression in the LS and NLS of patients with psoriasis in the (B) GSE53552 and (E) GSE13355 datasets. Receiver operating characteristic curves of TGM1 in the (C) GSE53552 and (F) GSE13355 datasets. NLS, non-lesional skin; LS, lesional skin; FC, fold change; AUC, area under the curve; TGM1, transglutaminase 1.

Article Snippet: Sections were incubated with a primary antibody against TGM1 (1: 100 cat. no. 12912-3-AP; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Biomarker Discovery, Expressing

Journal: Molecular Medicine Reports

Article Title: TGM1 as a novel signature gene in psoriasis identified by integrative bioinformatics and experimental validation

doi: 10.3892/mmr.2025.13755

Figure Lengend Snippet: TGM1 expression in psoriatic skin validated by IHC. (A) H&E staining of skin tissues of healthy controls and patients with psoriasis. (B) IHC of TGM1 protein in skin tissues of healthy controls and patients with psoriasis. (C) TGM1-positive cells in skin tissues of healthy controls and patients with psoriasis. ***P<0.001 vs. healthy control. IHC, immunohistochemistry; TGM1, transglutaminase 1.

Article Snippet: Sections were incubated with a primary antibody against TGM1 (1: 100 cat. no. 12912-3-AP; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Staining, Control, Immunohistochemistry

Journal: Molecular Medicine Reports

Article Title: TGM1 as a novel signature gene in psoriasis identified by integrative bioinformatics and experimental validation

doi: 10.3892/mmr.2025.13755

Figure Lengend Snippet: Correlation of TGM1 expression with infiltrating immune cells in psoriasis. (A) Expression of immune cells between LS and NLS. (B) Correlation between 22 types of infiltrating immune cells between LS and NLS. (C) Lollipop graph showing the correlation between TGM1 expression and 22 types of infiltrating immune cells in psoriasis. Scatter plots showing that (D) eosinophils, (E) activated dendritic cells, (F) follicular T helper cells and (G) resting mast cells were significantly correlated with TGM1 expression. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 vs. NLS. NLS, non-lesional skin; ns, not significant; TGM1, transglutaminase 1; NK, natural killer; abs, absolute; cor, correlation coefficient.

Article Snippet: Sections were incubated with a primary antibody against TGM1 (1: 100 cat. no. 12912-3-AP; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: TGM1 as a novel signature gene in psoriasis identified by integrative bioinformatics and experimental validation

doi: 10.3892/mmr.2025.13755

Figure Lengend Snippet: mRNA expression levels of inflammatory cytokines and differentiation markers of keratinocytes after TGM1 overexpression. TGM1 (A) mRNA and (B) protein expression following sh-TGM1 treatment. (C) mRNA expression levels of inflammatory factors IL-1α, IL-1β, IL-6 and IL-23. mRNA expression levels of (D) S100A8 and S100A9, as well as (E) K1, K6, K10, K16 and K17 after sh-TGM1 treatment. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001. Neg, negative control; S100, S100 calcium binding protein; K, keratin; TGM1, transglutaminase 1.

Article Snippet: Sections were incubated with a primary antibody against TGM1 (1: 100 cat. no. 12912-3-AP; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Over Expression, Negative Control, Binding Assay

Journal: Molecular Medicine Reports

Article Title: TGM1 as a novel signature gene in psoriasis identified by integrative bioinformatics and experimental validation

doi: 10.3892/mmr.2025.13755

Figure Lengend Snippet: mRNA expression levels of inflammatory cytokines and differentiation markers of keratinocytes after TGM1 knockdown. TGM1 (A) mRNA and (B) protein expression after si-TGM1 transfection. (C) mRNA expression levels of inflammatory factors IL-1α, IL-1β and IL-6. mRNA expression levels of (D) S100A8 and S100A9, as well as (E) K1, K6, K10, K16 and K17 following M5 stimulation after si-TGM1 transfection. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 vs. si-Neg. si, small interfering RNA; Neg, negative control; TGM1, transglutaminase 1; S100, S100 calcium binding protein; K, keratin.

Article Snippet: Sections were incubated with a primary antibody against TGM1 (1: 100 cat. no. 12912-3-AP; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Knockdown, Transfection, Small Interfering RNA, Negative Control, Binding Assay