Journal: Redox Biology

Article Title: Small molecule HCY-NBD stabilizes GSTM2 via cys174 sulfenylation to attenuate high glucose induced endothelial cell senescence and calcification

doi: 10.1016/j.redox.2026.104058

Figure Lengend Snippet: HCY-NBD binds to GSTM2 and stabilize its protein levels (A) Three-dimensional schematic diagram of the interaction between HCY-NBD and GSTM2. The yellow and gray structures represent the 3D conformation of GSTM2 protein, while HCY-NBD is depicted as a green stick model. Molecular docking simulations predict binding affinity between GSTM2 and HCY-NBD with a binding energy of −7.52 kcal/mol. (B) Two-dimensional interaction diagram between HCY-NBD and GSTM2. Green dashed lines indicate hydrogen bonds, light green dashed lines represent carbon-hydrogen bonds, and pink dashed lines denote hydrophobic interactions. (C) Binding kinetics between HCY-NBD and GSTM2 measured by Biacore T200. (D) GSTM2 enzymatic activity kinetics was measured in vitro following exposure to either DMSO solvent control or HCY-NBD. GS-DNB refers to the product of the GSTM2 enzymatic activity assay. (E) HUVEC cells were treated with increasing glucose concentrations (0, 10, 30, 60 mM) for 24 h, followed by quantification of GSTM2 mRNA levels via qPCR. (F) HUVECs were treated with HCY-NBD (0, 1, 3, 5 μM) for 24 h under HG (30 mM) conditions, the GSTM2 mRNA expression were analyzed by qPCR. (G – H) HUVECs were exposed to 0 or 5 μM HCY-NBD under either normal-glucose (5.5 mM) or HG (30 mM) conditions for 24 h, the GSTM2 protein levels were determined by Western blot. (I – J) HUVECs were treated with HCY-NBD (0, 1, 3, 5 μM) for 24 h under HG (30 mM) conditions, the GSTM2 protein levels were determined by Western blot. One-way ANOVA followed by Tukey's post-hoc test was performed, ns p > 0.05, ∗p < 0.05, ∗∗p < 0.01, n = 3.

Article Snippet: Briefly, 2 μg of GSTM2 (HY–P75152A, MCE) protein was incubated with DMSO or HCY-NBD (0.1, 0.5, 1, 3, 5 μM) on ice for 1 h. The enzymatic reaction was initiated by adding the reaction mixture containing 1 mM GSH and 1 mM CDNB in a 96-well UV-transparent microplate.

Techniques: Binding Assay, Activity Assay, In Vitro, Solvent, Control, Enzyme Activity Assay, Expressing, Western Blot

Journal: Redox Biology

Article Title: Small molecule HCY-NBD stabilizes GSTM2 via cys174 sulfenylation to attenuate high glucose induced endothelial cell senescence and calcification

doi: 10.1016/j.redox.2026.104058

Figure Lengend Snippet: HCY-NBD up-regulates GSTM2 sulfenylation and inhibits GSTM2 ubiquitination (A-B) HUVECs were pretreated with 80 μM HDX for 1h, then the cells were treated with HCY-NBD (0 or 5 μM) and HDX (0 or 80 μM) under HG (30 mM) conditions for 24 h. Protein levels of GSTM2 were analyzed by Western blot. (C – D) HUVECs were exposed to 0 or 5 μM HCY-NBD under either normal-glucose (5.5 mM) or HG (30 mM) conditions for 24 h, the sulfenylation levels of GSTM2 were measured by Western blot. (E – F) HUVECs were exposed to 0 or 5 μM HCY-NBD under either normal-glucose (5.5 mM) or HG (30 mM) conditions for 24 h, the ubiquitination levels of GSTM2 were determined by Western blot. (G – H) HUVECs were transfected with GFP-GSTM2, and HA-Ub (Wild-type, K48-only, or K63-only), exposed to designated treatments, and assessed for GFP-GSTM2 ubiquitination levels via Western blot in all experimental groups. (I – L) HUVECs were exposed to 0 or 5 μM HCY-NBD under either normal-glucose (5.5 mM) or HG (30 mM) conditions for 24 h, the glutathionylation levels of GSTM2 were determined by Western blot. One-way ANOVA followed by Tukey's post-hoc test was performed, ns p > 0.05, ∗p < 0.05, ∗∗p < 0.01, n = 3.

Article Snippet: Briefly, 2 μg of GSTM2 (HY–P75152A, MCE) protein was incubated with DMSO or HCY-NBD (0.1, 0.5, 1, 3, 5 μM) on ice for 1 h. The enzymatic reaction was initiated by adding the reaction mixture containing 1 mM GSH and 1 mM CDNB in a 96-well UV-transparent microplate.

Techniques: Ubiquitin Proteomics, Western Blot, Transfection

Journal: Redox Biology

Article Title: Small molecule HCY-NBD stabilizes GSTM2 via cys174 sulfenylation to attenuate high glucose induced endothelial cell senescence and calcification

doi: 10.1016/j.redox.2026.104058

Figure Lengend Snippet: Cys174 is a critical residue for HCY-NBD-mediated regulation of GSTM2 sulfenylation and non-lysine ubiquitination (A-B) HUVECs were transfected with wide type (WT) and four distinct cysteine-to-alanine mutants (C87A, C115A, C174A, and C87-115-174A) of GSTM2, exposed to designated treatments, and assessed for GSTM2 sulfenylation levels via Western blot in all experimental groups. (C) HEK-293T cells were transfected with a GSTM2-overexpressing plasmid for 48 h. Ubiquitination at Cys174 was detected via LC-MS/MS, with MS/MS analysis demonstrating a characteristic +114.043 Da mass shift (C [114.043]). Score, 131.0. (D – E) HUVECs were transfected with wide type (WT) and 174 cysteine-to-alanine(C174A) mutants of GSTM2, exposed to designated treatments, and assessed for GSTM2 ubiquitination levels via Western blot in all experimental groups. (F – G) HUVECs were transfected with GFP-GSTM2-WT, GFP-GSTM2-C174A, and HA-Ub (WT, K48-only), exposed to designated treatments, and assessed for GFP-GSTM2 ubiquitination levels via Western blot in all experimental groups. (H–K) HUVECs were transfected with GFP-GSTM2-WT or GFP-GSTM2-C174A 24 h, then treated with cycloheximide (2.5 μM) and designated treatments for the indicated times, followed by Western blotting. (L – M) HUVECs were transfected with wide type (WT) and 174 cysteine-to-alanine(C174A) mutants of GSTM2, exposed to designated treatments, and assessed for HA levels via Western blot in all experimental groups. H–K: Two-way ANOVA was performed, others: one-way ANOVA followed by Tukey's post-hoc test was performed, ns p > 0.05, ∗p < 0.05, ∗∗p < 0.01, n = 3.

Article Snippet: Briefly, 2 μg of GSTM2 (HY–P75152A, MCE) protein was incubated with DMSO or HCY-NBD (0.1, 0.5, 1, 3, 5 μM) on ice for 1 h. The enzymatic reaction was initiated by adding the reaction mixture containing 1 mM GSH and 1 mM CDNB in a 96-well UV-transparent microplate.

Techniques: Residue, Ubiquitin Proteomics, Transfection, Western Blot, Plasmid Preparation, Liquid Chromatography with Mass Spectroscopy, Tandem Mass Spectroscopy

Journal: Redox Biology

Article Title: Small molecule HCY-NBD stabilizes GSTM2 via cys174 sulfenylation to attenuate high glucose induced endothelial cell senescence and calcification

doi: 10.1016/j.redox.2026.104058

Figure Lengend Snippet: HCY-NBD inhibits high glucose-induced senescence of HUVECs (A-D) HUVECs were exposed to 0 or 5 μM HCY-NBD under either normal-glucose (5.5 mM) or HG (30 mM) conditions for 24 h, the protein levels of p21, p16 and p53 were determined by Western blot. (E – F) Cell cycle analysis by flow cytometry for HUVECs in different groups. (G – H) HUVECs were exposed to 0 or 5 μM HCY-NBD under either normal-glucose (5.5 mM) or HG (30 mM) conditions for 24 h, SA- β -gal staining was performed to observe senescence in HUVECs. (I – L) HUVECs were transfected with GSTM2 siRNA at a final concentration of 50 nM, while control groups were transfected with scramble siRNA at the same concentration. After 24 h, cells cultured under a HG (30 mM) model were treated with 0 or 5 μM HCY-NBD for 24 h. Protein levels of p21 were analyzed by Western blot and SA- β -gal staining was performed to observe senescence in HUVECs. (M – N) HUVECs were transfected with wide type (WT) and four distinct cysteine-to-alanine mutants (C87A, C115A, C174A, and C87-115-174A) of GSTM2, exposed to designated treatments, and assessed for protein levels of p21 via Western blot in all experimental groups. One-way ANOVA followed by Tukey's post-hoc test was performed, ns p > 0.05, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n = 3, n = 4(E–F).

Article Snippet: Briefly, 2 μg of GSTM2 (HY–P75152A, MCE) protein was incubated with DMSO or HCY-NBD (0.1, 0.5, 1, 3, 5 μM) on ice for 1 h. The enzymatic reaction was initiated by adding the reaction mixture containing 1 mM GSH and 1 mM CDNB in a 96-well UV-transparent microplate.

Techniques: Western Blot, Cell Cycle Assay, Flow Cytometry, Staining, Transfection, Concentration Assay, Control, Cell Culture

Journal: Redox Biology

Article Title: Small molecule HCY-NBD stabilizes GSTM2 via cys174 sulfenylation to attenuate high glucose induced endothelial cell senescence and calcification

doi: 10.1016/j.redox.2026.104058

Figure Lengend Snippet: HCY-NBD inhibits vascular senescence in db/db mice (A) Experimental Workflow Diagram. (B) HCY-NBD was administered intraperitoneally to db/db mice (0, 1, or 5 mg/kg per injection, every other day) over an 8-week period. Body weights were recorded weekly, n = 5. (C–H) Following 8 weeks of treatment, mice were humanely euthanized. Major organs (heart, liver, spleen, lungs, and kidneys) were excised and weighed using an analytical balance. Bar charts depict organ-to-body weight ratios, calculated as (organ weight/total body weight) × 100 %, n = 5. (I) Histopathological evaluation of heart, liver, spleen, lung, and kidney tissues was performed on H&E-stained sections to characterize microstructural organization and injury phenotypes, n = 4. (J – L) Thoracic aortic segments were subjected to en face immunofluorescence staining to assess p21 levels within CD31-positive endothelial cells and CD31 levels. Nuclei were counterstained with DAPI. n = 4. (M – N) Vascular senescence in thoracic aortas was detected using SA- β -gal staining, n = 3. (O–P) Protein expression levels of GSTM2 in thoracic aortas of db/db mice were detected via immunohistochemistry, n = 5. (Q – R) Immunofluorescence analysis detected protein expression levels of GSTM2 colocalized with CD31 in thoracic aortas of db/db mice, n = 4. One-way ANOVA followed by Tukey's post-hoc test was performed, ns p > 0.05, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Briefly, 2 μg of GSTM2 (HY–P75152A, MCE) protein was incubated with DMSO or HCY-NBD (0.1, 0.5, 1, 3, 5 μM) on ice for 1 h. The enzymatic reaction was initiated by adding the reaction mixture containing 1 mM GSH and 1 mM CDNB in a 96-well UV-transparent microplate.

Techniques: Injection, Mass Measurement, Staining, Immunofluorescence, Expressing, Immunohistochemistry

Journal: Redox Biology

Article Title: Small molecule HCY-NBD stabilizes GSTM2 via cys174 sulfenylation to attenuate high glucose induced endothelial cell senescence and calcification

doi: 10.1016/j.redox.2026.104058

Figure Lengend Snippet: Schematic presentation of the new approach to enhance endogenous GSTM2 level that inhibits high glucose-induced senescence and calcification in vascular endothelial cells.

Article Snippet: Briefly, 2 μg of GSTM2 (HY–P75152A, MCE) protein was incubated with DMSO or HCY-NBD (0.1, 0.5, 1, 3, 5 μM) on ice for 1 h. The enzymatic reaction was initiated by adding the reaction mixture containing 1 mM GSH and 1 mM CDNB in a 96-well UV-transparent microplate.

Techniques:

Journal: Cell Metabolism

Article Title: Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage

doi: 10.1016/j.cmet.2020.06.004

Figure Lengend Snippet: sEV-Ys Are Enriched in Proteins Related to Glutathione Conjugation and Ameliorate ROS Levels in Old Donor-Derived Fibroblasts (A) Analysis of a published proteomic analysis ( Borghesan et al., 2019 ) on sEVs isolated from iC fibroblasts and sEVs derived from cells undergoing senescence (iRAS and by etoposide treatment). We found 55 proteins enriched in sEVs derived from iC in comparison with sEVs from iRAS and etoposide. (B and C) STRING (B) and Reactome pathway analysis (C) for the 55 proteins enriched in sEVs from iC highlight the glutathione metabolism pathway where GSTM2 is highlighted. (D) Immunoblotting for GSTM2 and ALIX in sEVs isolated from young, old, and HGPS fibroblasts loading the same number of sEVs (3 × 10 9 ). sEVs isolated from all four cell lines are shown. (E) Densitometry quantification for GSTM2 immunoblotting versus ALIX. t test analyses were performed. ∗p < 0.05; ∗∗p < 0.01. (F) sEVs were subjected to density gradient by Optiprep and different fractions were immunoblotted for GSTM2 and TSG101 showing the presence of sEVs containing GSTM2. (G) IF quantification for ROS by 8-oxodG staining in 4 old fibroblasts treated with the CM and other fractions derived from young cells. t test analyses were performed. ∗∗p < 0.01; ns, non-significant. (H) Representative pictures for 8-oxodG staining in GM05565 (old cell line) treated with sEV-Ys and sEV-Os or with FBS 10%. (I) IF quantification for p-γH2AX staining in 4 old fibroblasts treated with young donor CM and fractions. t test analyses were performed. ∗∗p < 0.01; ∗∗∗p < 0.001; ns, non-significant. (J) Pictures of p-γH2AX in AG16086 (old cell line) basal levels or treated with sEV-Ys or sEV-Os. (H and J) Scale bar, 50 μm. (G and I) FBS 10% is shown as negative control. All data represent mean ± SEM in 4 different cells lines derived from old donors. See also Figure S3 .

Article Snippet: GSTM2 Protein, Human, Recombinant (His Tag) , Sino Biological , 12042-H07E.

Techniques: Conjugation Assay, Derivative Assay, Isolation, Western Blot, Staining, Negative Control

Journal: Cell Metabolism

Article Title: Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage

doi: 10.1016/j.cmet.2020.06.004

Figure Lengend Snippet: GST Activity and GSH Levels Are Important in Mediating sEV-Y Rejuvenation in Old Donors (A) sEVs isolated from 4 different young donors have independent GST activity. sEVs from old donors and their respective SF fractions from young and old donors do not present GST activity. t test analysis was performed. ∗∗p < 0.01. (B) GST activity was determined in old fibroblasts treated with sEVs from either young or old donors. FBS 10% was used as a control. Data show the mean ± SEM of 4 different donor cells. t test analysis was performed. ∗∗∗p < 0.001; ns, non-significant. (C) Ratio of GSH/GSSG in old cells treated with sEVs and different concentrations (20 or 40 μM) of BSO (buthionine sulphoximine), which prevent de novo GSH synthesis. The increase in GSH/GSSG levels when old cells are treated with sEV-Ys is prevented after BSO treatment. ∗p < 0.05; ∗∗p < 0.01; ns, non-significant. (D) Relative cell number shows an increase in proliferation in old cells treated with sEV-Ys, which is prevented by GSH inhibition (BSO). ∗∗p < 0.01. (E) SA-β-Gal activity downregulation by sEV-Ys is prevented by 20 μM BSO treatment. ∗p < 0.05; ∗∗p < 0.01. (F and G) iC or iRAS HFFF2 cells were treated with sEVs derived from iC or iRAS ectopically expressing myc- Gstm2 or empty vector. The mean ± SEM from three independent experiments is shown. (F) SA-β-Gal activity was quantified and (G) representative images are shown. ∗∗p < 0.01; ns, non-significant. (H) Diagram of the protocol followed to transfect recombinant GSTM2 (rGSTM2) into old sEVs. (I) Four old donor cells were treated with sEVs isolated from old and young donors transfected with either IgG or rGSTM2 (rGSTM2-sEV). rGSTM2 was used on old donor cells on its own as a positive control. SA-β-Gal activity quantification and representative pictures are shown. Quantification represents the mean ± SEM of 4 different donor cell lines. ∗∗p < 0.01; ns, non-significant. See also Figure S4 .

Article Snippet: GSTM2 Protein, Human, Recombinant (His Tag) , Sino Biological , 12042-H07E.

Techniques: Activity Assay, Isolation, Inhibition, Derivative Assay, Expressing, Plasmid Preparation, Recombinant, Transfection, Positive Control

Journal: Cell Metabolism

Article Title: Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage

doi: 10.1016/j.cmet.2020.06.004

Figure Lengend Snippet: sEV-Ys Prevent ROS Accumulation and Increase the Levels of GSH in Old Mice (A) The levels of oxidative stress (ROS) were measured in liver and serum from young mice (n = 3 mice) and old mice treated or not with sEV-Ys (n = 5 old mice per condition). Data represent mean ± SEM. t test analysis was performed. (B) Representative immunoblot for the expression of GSTM2 in the liver in young and old treated or untreated mice. GAPDH was used as loading control and quantification of 5 mice per condition is shown in graph. (C) Measurement of GST activity in the liver and serum from different young, old sEV-Y-treated, or non-treated mice (n = 3 young and n = 5 per condition in old mice). Data represent mean ± SEM. t test was performed. (D) qPCR analysis for different transcripts related to the antioxidant pathway in the liver from young, old sEV-Y-treated, or untreated old mice (n = 3 young and n = 5 per condition in old mice). Mann-Whitney test was performed. See also Figure S6 .

Article Snippet: GSTM2 Protein, Human, Recombinant (His Tag) , Sino Biological , 12042-H07E.

Techniques: Western Blot, Expressing, Activity Assay, MANN-WHITNEY

Journal: Cell Metabolism

Article Title: Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage

doi: 10.1016/j.cmet.2020.06.004

Figure Lengend Snippet: sEV-Ys Ameliorate Lipid Peroxidation In Vitro and In Vivo (A and B) MDA levels in (A) liver and (B) serum from young and old mice treated with sEV-Ys or not. Data represent the mean ± SEM of 3–5 mice. t test analysis was performed. (C) MDA quantification in iRAS cells treated with sEVs from iC or iRAS expressing myc- Gstm2 or not. Data show the mean ± SEM of 3 independent experiments. ∗p < 0.05; ns, non-significant. (D) Quantification of MDA in old donor fibroblasts incubated with sEVs isolated from either young or old donors treated with 20 μM BSO. Graphs represents the mean ± SEM of 4 old donor fibroblasts. ∗∗∗p < 0.001; ns, non-significant. (E) Growth curve for young HFFF2 fibroblasts treated with increasing concentrations of 4-HNE (1.25, 2.5, and 5 μM). iRAS were used as a positive control. Data represent the mean ± SEM of 3 independent experiments. Two-way ANOVA was performed. ∗∗∗p < 0.001. (F) IF analyses of biomarkers of senescence (SA-β-Gal and p16 INK4A ) in 4-HNE-treated young fibroblasts with different siRNA targeting a variety of pathways that regulate senescence. 50 nM siRNA targeting p16 (sip16), p53 (sip53), NF-κB (sip65), and C/EBPβ (siCEBP) was used. Scr is a non-targeting control. ∗∗∗p < 0.001. (G) MDA levels in 4-HNE-treated young HFFF2 cells incubated or not with sEV-Ys. ∗∗∗p < 0.001. (H) Quantification of SA-β-Gal in HFFF2 cells treated with 4-HNE and different inhibitors targeting IKK (NF-κB pathway; 20 μM CAY10576) or ROS (100 nM NAC). sEV-Ys were used as a positive control. ∗p < 0.05. See also Figure S7 .

Article Snippet: GSTM2 Protein, Human, Recombinant (His Tag) , Sino Biological , 12042-H07E.

Techniques: In Vitro, In Vivo, Expressing, Incubation, Isolation, Positive Control

Journal: Cell Metabolism

Article Title: Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage

doi: 10.1016/j.cmet.2020.06.004

Figure Lengend Snippet:

Article Snippet: GSTM2 Protein, Human, Recombinant (His Tag) , Sino Biological , 12042-H07E.

Techniques: Recombinant, Immunofluorescence, Glutathione S-Transferase Assay, Peroxidation Assay, Fluorescence, TBARS Assay, Software, Functional Assay, Protein Concentration, Enzyme-linked Immunosorbent Assay

Journal: The American journal of pathology

Article Title: Pseudogene-Derived Long Noncoding RNAs GSTM3P1/Gstm2-ps1 Exacerbate Sepsis-Associated Acute Kidney Injury by Suppressing Their Parent Gene Translation

doi: 10.1016/j.ajpath.2026.01.004

Figure Lengend Snippet: GSTM3P1/Gstm2-ps1 are transiently induced in kidney proximal tubular cells in sepsis-associated acute kidney injury models. BUMPT and HK-2 cells were treated with 100 μg/mL lipopolysaccharide (LPS) for 3, 6, and 12 hours. Male C57BL/6J mice were subjected to an i.p. injection of 10 mg/kg LPS or cecal ligation and puncture (CLP) surgery, and kidney samples were collected at 3, 6, and 9 hours after injury. A—D: Quantitative RT-PCR analysis of Gstm2-ps1 or GSTM3P1 expression. A: Gstm2-ps1 in BUMPT. B: GSTM3P1 in HK2. C: Gstm2-ps1 in LPS-treated mouse kidney. D: Gstm2-ps1 in mouse kidney with CLP. A—D: The statistical difference was determined using one-way analysis of variance with Tukey multiple comparison. E and F: Representative images of Gstm2-ps1 in kidney tissue assessed by in situ hybridization and costained with fluorescein isothiocyanate—labeled Lotus tetragonolobus lectin (LTL). E: Mouse kidneys with or without LPS treatment for 3 hours. F: Mouse kidneys with or without CLP injury for 3 hours. n 5 ( A—D ); n 6 ( E and F ). ** P < 0.01, *** P < 0.001. Scale bar 100 μm ( E and F ). NS, nonsignificant.

Article Snippet: Human GSTM3 or murine Gstm2 open reading frames were subcloned from commercially available plasmids (RC201013 or MR202469; Origene, Rockville, MD), respectively.

Techniques: Injection, Ligation, Quantitative RT-PCR, Expressing, Comparison, In Situ Hybridization, Labeling

Journal: The American journal of pathology

Article Title: Pseudogene-Derived Long Noncoding RNAs GSTM3P1/Gstm2-ps1 Exacerbate Sepsis-Associated Acute Kidney Injury by Suppressing Their Parent Gene Translation

doi: 10.1016/j.ajpath.2026.01.004

Figure Lengend Snippet: GSTM3P1/Gstm2-ps1 exacerbate lipopolysaccharide (LPS)—induced apoptosis in cultured proximal tubular cells. RPTC cells with or without GSTM3P1 stable overexpression, BUMPT cells with or without Gstm2-ps1 stable overexpression, and wild-type (WT) or Gstm2-ps1 knockout (KO) primary proximal tubular cells were treated with 100 μg/mL LPS for 19 hours. A—C: Quantitative RT-PCR analysis confirming the expression of GSTM3P1 or Gstm2-ps1. A: RPTC. B: BUMPT. C: Primary proximal tubular cells. D—F: Cell viability assessed by MTT assay following 19 hours of LPS treatment. D: RPTC. E: BUMPT. F: Primary proximal tubular cells. A—F: The statistical difference was determined by unpaired t -test. G—I: Representative immunoblots of full-length caspase 3 and cleaved caspase 3 (C-caspase-3) with β-actin as the loading control (Ctrl; top panels ), and the densitometry analysis of C-caspase-3 normalized to β-actin ( bottom panels ). G: RPTC. H: BUMPT. I: Primary proximal tubular cells. G—I: The statistical difference was determined by two-way analysis of variance with Tukey multiple comparison. n =3 ( A , G , and H ); n = 4 ( B , C , and I ); n = 6 ( D—F ). * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Human GSTM3 or murine Gstm2 open reading frames were subcloned from commercially available plasmids (RC201013 or MR202469; Origene, Rockville, MD), respectively.

Techniques: Cell Culture, Over Expression, Knock-Out, Quantitative RT-PCR, Expressing, MTT Assay, Western Blot, Control, Comparison

Journal: The American journal of pathology

Article Title: Pseudogene-Derived Long Noncoding RNAs GSTM3P1/Gstm2-ps1 Exacerbate Sepsis-Associated Acute Kidney Injury by Suppressing Their Parent Gene Translation

doi: 10.1016/j.ajpath.2026.01.004

Figure Lengend Snippet: GSTM3P1/Gstm2-ps1 overexpression induces oxidative stress in cultured proximal tubular cells. RPTC cells with or without GSTM3P1 stable overexpression and BUMPT cells with or without Gstm2-ps1 stable overexpression were treated with 100 μg/mL lipopolysaccharide (LPS) for 19 hours. A and B: Oxidative stress levels were assessed by measuring the reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio. A: RPTC. B: BUMPT. The statistical difference was determined by two-way analysis of variance with Tukey multiple comparison. C: Representative images of 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA)—stained RPTC and phase-contrast images. n = 6 ( A—C ). * P < 0.05, ** P < 0.01, and *** P < 0.001. Scale bars = 200 μm ( C ).

Article Snippet: Human GSTM3 or murine Gstm2 open reading frames were subcloned from commercially available plasmids (RC201013 or MR202469; Origene, Rockville, MD), respectively.

Techniques: Over Expression, Cell Culture, Comparison, Staining

Journal: The American journal of pathology

Article Title: Pseudogene-Derived Long Noncoding RNAs GSTM3P1/Gstm2-ps1 Exacerbate Sepsis-Associated Acute Kidney Injury by Suppressing Their Parent Gene Translation

doi: 10.1016/j.ajpath.2026.01.004

Figure Lengend Snippet: Proximal tubular—specific Gstm2-ps1 knockout protects mice from lipopolysaccharide (LPS)—induced sepsis acute kidney injury. Proximal tubular—specific Gstm2-ps1 knockout (KO) mice and their wild-type (WT) littermates were treated with one i.p. injection of 10 mg/kg LPS and sacrificed after 19 hours. The control (Ctrl) mice were given an equal volume of phosphate-buffered saline. A: Blood urea nitrogen (BUN) levels. B: Serum creatinine levels. C: Quantification of apoptotic cells in kidneys by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. A—C: The statistical difference was determined by two-way analysis of variance with Tukey multiple comparison. D: Representative images of TUNEL staining. E: Representative images of neutrophil gelatinase-associated lipocalin (NGAL) immunohistochemical staining. F: Representative images of reactive oxygen species (ROS) levels detected by 2′,7′-dichlorodihydrofluorescein diacetate staining. G: Representative immunoblots of kidney injury molecule-1 (KIM-1) and the densitometry analysis of KIM-1 normalized to the internal loading control glyceraldehyde-3-phosphate dehydrogenase. Data are presented as means ± SD ( A—C and G ). n = 6 per group ( A , B , E , and F ); n = 5 per group ( C ); n = 4 per group ( G ). ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Scale bars: 50 μm ( D and E ); 100 μm ( F ).

Article Snippet: Human GSTM3 or murine Gstm2 open reading frames were subcloned from commercially available plasmids (RC201013 or MR202469; Origene, Rockville, MD), respectively.

Techniques: Knock-Out, Injection, Control, Saline, End Labeling, TUNEL Assay, Staining, Comparison, Immunohistochemical staining, Western Blot

Journal: The American journal of pathology

Article Title: Pseudogene-Derived Long Noncoding RNAs GSTM3P1/Gstm2-ps1 Exacerbate Sepsis-Associated Acute Kidney Injury by Suppressing Their Parent Gene Translation

doi: 10.1016/j.ajpath.2026.01.004

Figure Lengend Snippet: GSTM3P1/Gstm2-ps1 down-regulate the expression of their parent gene GSTM3/GSTM2. A—F: RPTC cells ( A and D ) or BUMPT cells ( B and E ) were overexpressed with or without GSTM3P1 or Gstm2-ps1, and C57BL/6J mouse kidneys ( C and F ) were treated with 10 mg/kg lipopolysaccharide (LPS) for 3, 6, and 9 hours. A—C: Representative immunoblots of GSTM3 and GSTM2, with β-actin or cyclophilin B as the internal loading marker. D—F: The densitometric analysis of GSTM3 and GSTM2 protein normalized to the internal loading control. The statistical difference was determined by unpaired t -test ( D and E ) or one-way analysis of variance with Tukey multiple comparison ( F ). G: Representative immunofluorescence images of GSTM2 with fluorescein isothiocyanate— Lotus tetragonolobus lectin (LTL) costaining. Gstm2-ps1 knockout (KO) and wild-type (WT) mice were treated with/without 10 mg/kg LPS for 3 hours. n = 6 for cell samples ( D—F ); n = 4 for kidney samples ( D—F ); n = 6 ( G ). ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Scale bar = 100 μm ( G ). NS, nonsignificant.

Article Snippet: Human GSTM3 or murine Gstm2 open reading frames were subcloned from commercially available plasmids (RC201013 or MR202469; Origene, Rockville, MD), respectively.

Techniques: Expressing, Western Blot, Marker, Control, Comparison, Immunofluorescence, Knock-Out

Journal: The American journal of pathology

Article Title: Pseudogene-Derived Long Noncoding RNAs GSTM3P1/Gstm2-ps1 Exacerbate Sepsis-Associated Acute Kidney Injury by Suppressing Their Parent Gene Translation

doi: 10.1016/j.ajpath.2026.01.004

Figure Lengend Snippet: GSTM2/GSTM3 protect cultured renal proximal tubular cell from lipopolysaccharide (LPS)—induced apoptosis. RPTC cells with or without GSTM3 overexpression, and BUMPT cells with or without GSTM2 overexpression, were treated with 100 μg/mL LPS. A and B: Immunoblot analysis confirming the overexpression efficiency of GSTM3 and GSTM2. C and D: Cell viability assessed using the MTT assay following 17 hours of LPS treatment. A—D: The statistical difference was determined by unpaired t -test. E and F: Representative immunoblots ( top panels ) and densitometric analysis ( bottom panels ) of cleaved caspase-3 (C-caspase-3) protein levels. E: RPTC. F: BUMPT. E and F: The statistical difference was determined by one-way analysis of variance with Tukey multiple comparison. n = 3 ( A and B ); n = 6 ( C and D ); n = 4 ( E and F ). ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Ctrl, control.

Article Snippet: Human GSTM3 or murine Gstm2 open reading frames were subcloned from commercially available plasmids (RC201013 or MR202469; Origene, Rockville, MD), respectively.

Techniques: Cell Culture, Over Expression, Western Blot, MTT Assay, Comparison, Control

Journal: The American journal of pathology

Article Title: Pseudogene-Derived Long Noncoding RNAs GSTM3P1/Gstm2-ps1 Exacerbate Sepsis-Associated Acute Kidney Injury by Suppressing Their Parent Gene Translation

doi: 10.1016/j.ajpath.2026.01.004

Figure Lengend Snippet: GSTM3P1/Gstm2-ps1 may down-regulate GSTM3/GSTM2 protein level through RNA translation inhibition. A: Quantitative RT-PCR (RT-qPCR) of GSTM3 in RPTC cells with or without GSTM3P1 overexpression. The unpaired t -test was used. B: RT-qPCR of GSTM2 in BUMPT cells with or without Gstm2-ps1 overexpression. The unpaired t -test was used. C: RT-qPCR of GSTM2 in mouse kidneys with or without 10 mg/kg lipopolysaccharide (LPS) treatment . The one-way analysis of variance (ANOVA) with Tukey multiple comparison was used. D and E: BUMPT cell lysates with Gstm2-ps1 overexpression or empty vector transfection (pcDNA) were subjected to RNA pull-down assay with Gstm2-ps1 probes. The input lysates (Input) and pull-down products (pull-down) were examined by immunoblotting of human antigen R (HuR), with cyclophilin B as the internal loading control of input lysates. D: Representative immunoblots of HuR and cyclophilin B. E: The densitometry analysis of HuR immunoblotting in pull-down products normalized by its level in the input. The unpaired t -test was used. F—H: BUMPT cells were overexpressed with HuR or Gstm2-ps1. F: Representative immunoblots to confirm the overexpression of HuR with cyclophilin B as the internal loading control. The upper band is the overexpressed HuR. The lower band is the endogenous HuR. G: Representative immunoblots and densitometry analysis of GSTM2 in cells with or without Gstm2-ps1 and HuR overexpression. β-Actin was used as the internal loading control. The one-way ANOVA with Tukey multiple comparison was used. H: The cell viability assessed by MTT assay. The one-way ANOVA with Tukey multiple comparison was used. n = 5 ( A and B ); n = 4 ( C and E—G ); n = 6 ( H ). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. NS, nonsignificant.

Article Snippet: Human GSTM3 or murine Gstm2 open reading frames were subcloned from commercially available plasmids (RC201013 or MR202469; Origene, Rockville, MD), respectively.

Techniques: Inhibition, Quantitative RT-PCR, Over Expression, Comparison, Plasmid Preparation, Transfection, Pull Down Assay, Western Blot, Control, MTT Assay

Journal: Psychoneuroendocrinology

Article Title: Blood-Based Gene-Expression Biomarkers of Post-Traumatic Stress Disorder among Deployed Marines: A Pilot Study

doi: 10.1016/j.psyneuen.2014.09.024

Figure Lengend Snippet: Genes Significantly Dysregulated ( p <0.01) in Peripheral Blood Mononuclear Cells from the Full Sample of PTSD Cases at Post-Deployment and Used in Predictive SVM Classifiers *

Article Snippet: # Transcripts in bold comprised the optimal 2-probe SVM classifier of PTSD status identified by training and testing in independent samples. caption a8 Genes Significantly Dysregulated ( p <0.01) in Peripheral Blood Mononuclear Cells from a Subset of PTSD Cases at Post-Deployment and Used in Predictive SVM Classifiers * table ft1 table-wrap mode="anchored" t5 caption a7 Assay ID Gene GADPH -normalized Average 2 -ΔCt GADPH -normalized p -values HPRT1 -normalized Average 2 -ΔCt HPRT1 -normalized p -values PTSD Control PTSD Control Hs01683722_gH GSTM1 0.367±0.741 1.138±1.529 0.031* 0.685±1.367 1.251±1.139 0.128 Hs03044640_gH GSTM2 0.732±0.596 1.114±0.655 0.036* 0.300±0.184 0.531±0.262 0.001* Hs00180203_m1 CUL2 1.000±0.323 1.223±0.612 0.114 0.768±0.250 0.872±0.350 0.236 Hs00211676_m1 DYNC1LI1 0.277±0.261 0.333±0.204 0.401 0.441±0.434 0.466±0.243 0.803 Hs00948075_m1 HUWE1 0.333±0.204 0.586±0.570 0.192 0.144±0.105 0.208±0.215 0.184 Hs00277883_m1 LARP7 0.674±0.400 0.711±0.272 0.709 0.689±0.

Techniques: Diagnostic Assay, Binding Assay, Expressing, Coagulation

Journal: Psychoneuroendocrinology

Article Title: Blood-Based Gene-Expression Biomarkers of Post-Traumatic Stress Disorder among Deployed Marines: A Pilot Study

doi: 10.1016/j.psyneuen.2014.09.024

Figure Lengend Snippet: Genes Significantly Dysregulated ( p <0.01) in Peripheral Blood Mononuclear Cells from a Subset of PTSD Cases at Post-Deployment and Used in Predictive SVM Classifiers *

Article Snippet: # Transcripts in bold comprised the optimal 2-probe SVM classifier of PTSD status identified by training and testing in independent samples. caption a8 Genes Significantly Dysregulated ( p <0.01) in Peripheral Blood Mononuclear Cells from a Subset of PTSD Cases at Post-Deployment and Used in Predictive SVM Classifiers * table ft1 table-wrap mode="anchored" t5 caption a7 Assay ID Gene GADPH -normalized Average 2 -ΔCt GADPH -normalized p -values HPRT1 -normalized Average 2 -ΔCt HPRT1 -normalized p -values PTSD Control PTSD Control Hs01683722_gH GSTM1 0.367±0.741 1.138±1.529 0.031* 0.685±1.367 1.251±1.139 0.128 Hs03044640_gH GSTM2 0.732±0.596 1.114±0.655 0.036* 0.300±0.184 0.531±0.262 0.001* Hs00180203_m1 CUL2 1.000±0.323 1.223±0.612 0.114 0.768±0.250 0.872±0.350 0.236 Hs00211676_m1 DYNC1LI1 0.277±0.261 0.333±0.204 0.401 0.441±0.434 0.466±0.243 0.803 Hs00948075_m1 HUWE1 0.333±0.204 0.586±0.570 0.192 0.144±0.105 0.208±0.215 0.184 Hs00277883_m1 LARP7 0.674±0.400 0.711±0.272 0.709 0.689±0.

Techniques: Diagnostic Assay, Variant Assay, RNA Binding Assay, Binding Assay

Journal: Psychoneuroendocrinology

Article Title: Blood-Based Gene-Expression Biomarkers of Post-Traumatic Stress Disorder among Deployed Marines: A Pilot Study

doi: 10.1016/j.psyneuen.2014.09.024

Figure Lengend Snippet: Microarray-derived expression levels (ordinate) of summarized exon probesets reflecting whole-transcript expression levels (abscissa) of glutathione s-transferase mu 1 (GSTM1) and glutathione s-transferase mu 2 (GSTM2) in peripheral blood mononuclear cells from PTSD cases (n=25, red) and comparison subjects (n=25, blue). These transcripts were notably down-regulated among PTSD cases within the full sample (fold changes -1.58 and -2.00, respectively) and were identified as the sole components of the optimal performing SVM classifier of diagnostic status, which achieved 80% accuracy in the test subset (n=10; 4 of 5 cases correctly identified).

Article Snippet: # Transcripts in bold comprised the optimal 2-probe SVM classifier of PTSD status identified by training and testing in independent samples. caption a8 Genes Significantly Dysregulated ( p <0.01) in Peripheral Blood Mononuclear Cells from a Subset of PTSD Cases at Post-Deployment and Used in Predictive SVM Classifiers * table ft1 table-wrap mode="anchored" t5 caption a7 Assay ID Gene GADPH -normalized Average 2 -ΔCt GADPH -normalized p -values HPRT1 -normalized Average 2 -ΔCt HPRT1 -normalized p -values PTSD Control PTSD Control Hs01683722_gH GSTM1 0.367±0.741 1.138±1.529 0.031* 0.685±1.367 1.251±1.139 0.128 Hs03044640_gH GSTM2 0.732±0.596 1.114±0.655 0.036* 0.300±0.184 0.531±0.262 0.001* Hs00180203_m1 CUL2 1.000±0.323 1.223±0.612 0.114 0.768±0.250 0.872±0.350 0.236 Hs00211676_m1 DYNC1LI1 0.277±0.261 0.333±0.204 0.401 0.441±0.434 0.466±0.243 0.803 Hs00948075_m1 HUWE1 0.333±0.204 0.586±0.570 0.192 0.144±0.105 0.208±0.215 0.184 Hs00277883_m1 LARP7 0.674±0.400 0.711±0.272 0.709 0.689±0.

Techniques: Microarray, Derivative Assay, Expressing, Comparison, Diagnostic Assay

Journal: Psychoneuroendocrinology

Article Title: Blood-Based Gene-Expression Biomarkers of Post-Traumatic Stress Disorder among Deployed Marines: A Pilot Study

doi: 10.1016/j.psyneuen.2014.09.024

Figure Lengend Snippet: QRT-PCR Validation of Gene and Exon Expression in Full Sample of PTSD Cases and Comparison Subjects.

Article Snippet: # Transcripts in bold comprised the optimal 2-probe SVM classifier of PTSD status identified by training and testing in independent samples. caption a8 Genes Significantly Dysregulated ( p <0.01) in Peripheral Blood Mononuclear Cells from a Subset of PTSD Cases at Post-Deployment and Used in Predictive SVM Classifiers * table ft1 table-wrap mode="anchored" t5 caption a7 Assay ID Gene GADPH -normalized Average 2 -ΔCt GADPH -normalized p -values HPRT1 -normalized Average 2 -ΔCt HPRT1 -normalized p -values PTSD Control PTSD Control Hs01683722_gH GSTM1 0.367±0.741 1.138±1.529 0.031* 0.685±1.367 1.251±1.139 0.128 Hs03044640_gH GSTM2 0.732±0.596 1.114±0.655 0.036* 0.300±0.184 0.531±0.262 0.001* Hs00180203_m1 CUL2 1.000±0.323 1.223±0.612 0.114 0.768±0.250 0.872±0.350 0.236 Hs00211676_m1 DYNC1LI1 0.277±0.261 0.333±0.204 0.401 0.441±0.434 0.466±0.243 0.803 Hs00948075_m1 HUWE1 0.333±0.204 0.586±0.570 0.192 0.144±0.105 0.208±0.215 0.184 Hs00277883_m1 LARP7 0.674±0.400 0.711±0.272 0.709 0.689±0.

Techniques: Biomarker Discovery, Expressing, Comparison, Control

Journal: PLoS ONE

Article Title: Chronic Activation of Hepatic Nrf2 Has No Major Effect on Fatty Acid and Glucose Metabolism in Adult Mice

doi: 10.1371/journal.pone.0166110

Figure Lengend Snippet: Canonical pathways regulated by both siKeap1-1* and siKeap1-2*.

Article Snippet: Following primer sets were used: Keap1 (Mm00497268_m1), NAD(P)H dehydrogenase, quinone 1 (Nqo1, Mm00500821-m1), glutamatecysteine ligase, catalytic subunit (Gclc, Mm00802655-m1), glutathione reductase (Gsr, Mm00439154_m1), NF-E2-related factor 2 (Nrf2, Mm00477784_m1), glutathione S-transferase A2 (Gsta2, Mm03019257_g1), Gsta4 (Mm004974803_m1), thioredoxin reductase 1 (Txnrd1, Mm00443675_m1), glutathione S-transferase mu 1 (Gstm1, Mm00833915_g1), Gstm2 (Mm00725711_s1), Gstm3 (Mm00833923_m1), Gstm4 (Mm00728197_s1), sulfiredoxin 1 (Srxn1, Mm00769566_m1), microsomal glutathione S-transferase (Mgst3, Mm00787806_s1), and glutathione peroxidase 2 (Gpx2, Mm00850074_g1), glyceraldehyde-3-phosphate dehydrogenase (Gapdh, Mm03302249_g1) and ribosomal protein L37a (Rpl37a, Mm01546394_s1) as TaqMan® Gene Expression Assay (Thermo Scientific).

Techniques: