Journal: International Dental Journal

Article Title: Single-Cell and Machine Learning Analysis Reveal Novel Inflammatory Macrophage Subtypes and Biomarkers in Periodontitis

doi: 10.1016/j.identj.2025.103983

Figure Lengend Snippet: Construction and validation of a PMRGs diagnostic model. A, Violin plot depicting the PMRGs score levels in periodontal tissues of patients with PD ( n = 183) and healthy controls ( n = 64). Data are presented as means ± SD, **** P < .0001 vs control. B-F, ROC curves for (B) CBX4, (C) CXCR4, (D) MBP, (E) TXN and (F) ATF3 for external dataset validation. G, Identification of the optimal model using the average AUC from 101 diagnostic models across all training and test datasets. H-I, Leave-one-out cross-validation (LOOCV) framework was used to generate 101 prediction models, and the C-index was calculated for each model on both training and validation datasets. J, ROC curve for the PMRGs associated with PD-MΦ.

Article Snippet: Equal amounts of protein (30 μg/sample) were resolved by SDS-PAGE, transferred onto PVDF membranes and incubated overnight with primary antibodies specific for ATF3 (CST, #18665, 1:1500), CXCR4 (Abcam, ab181020, 1:1000), TXN (CST, #14907, 1:800), CBX3 (Abcam, ab217999, 1:1500), MBP (Abcam, ab11159, 1:800) and β-actin.

Techniques: Biomarker Discovery, Diagnostic Assay, Control

Journal: International Dental Journal

Article Title: Single-Cell and Machine Learning Analysis Reveal Novel Inflammatory Macrophage Subtypes and Biomarkers in Periodontitis

doi: 10.1016/j.identj.2025.103983

Figure Lengend Snippet: Expression levels of hub genes in periodontitis macrophages. A-G, The protein expression of ATF3, CXCR4, TXN, CBX3 and MBP. E-K, The mRNA expression of ATF3, CXCR4, TXN, CBX3 and MBP. Values represent the mean ± SD; ** P < .01, *** P < .001.

Article Snippet: Equal amounts of protein (30 μg/sample) were resolved by SDS-PAGE, transferred onto PVDF membranes and incubated overnight with primary antibodies specific for ATF3 (CST, #18665, 1:1500), CXCR4 (Abcam, ab181020, 1:1000), TXN (CST, #14907, 1:800), CBX3 (Abcam, ab217999, 1:1500), MBP (Abcam, ab11159, 1:800) and β-actin.

Techniques: Expressing

Journal: Life sciences

Article Title: Chronic administration of the hydrogen sulfide prodrug SG1002 partially protects against erectile dysfunction resulting from long-term androgen deprivation

doi: 10.1016/j.lfs.2025.123976

Figure Lengend Snippet: The expression of proteins related to the thioredoxin system in mouse corpus cavernosum measured by western blot. Mice underwent sham surgery (Sham) or were surgically castrated (Cast). Two groups of castrated mice were treated with low-dose (CLS) or high-dose (CHS) SG1002. Proteins include thioredoxin 1 (Trx1), thioredoxin 2 (Trx2), thioredoxin-interacting protein (Txnip), peroxiredoxin 3 (Prdx3), peroxiredoxin 5 (Prdx5). Protein expression was normalized to GAPDH. Values represent means ± SEM for n = 12 animals per group. * p < 0.05 compared to Sham.

Article Snippet: Primary antibodies were obtained from Cell Signaling Technologies (CST; Danvers, MA, USA), or Protein Tech (PT; Rosemount, IL, USA) and used at the following dilutions: glutamate-cysteine ligase (Gclc, PT #12601–1-AP, 1:1000), optic atrophy type 1 (Opa1, CST #80471, 1:1000), peroxiredoxin 3 (Prdx3, PT #10664–1-AP, 1:1000), peroxiredoxin 5 (Prdx5, PT #17724–1-AP, 1:1000), mitofusin 1 (Mfn1, PT #13798–1-AP, 1:1000), mitofusin 2 (Mfn2, PT #12186–1-AP, 1:1000), NAD(P)H dehydrogenase quinone 1 (Nqo1, CST #62262, 1:1000), thioredoxin 1 (Trx1, CST #2298, 1:1000), thioredoxin-interacting protein (Txnip, PT #18243–1-AP, 1:1000), thioredoxin 2 (Trx2, CST #14907, 1:1000), dynamin-related protein 1 (Drp1, CST #8570, 1:1000), superoxide dismutase 1 (Sod1, PT #10269–1-AP, 1:1000), superoxide dismutase 2 (Sod2, CST #13141, 1:2000), superoxide dismutase 3 (Sod3, Santa Cruz Biotechnology #sc-271170, 1:1000) autophagy-related protein (Atg7, CST #8558, 1:1000), mitochondrial fission 1 (Fis1, PT #10956–1-AP, 1:1000), phosphorylated (Ser 403 ) Sequestosome-1 (P-Sqstm1/p62, CST #39786, 1:1000), sequestosome-1 (Sqstm1/p62, CST #39749, 1:1000), heme oxygenase 1 (HO-1, PT #10701–1-AP, 1:1000), light chain 3B (LC3B, CST #2775, 1:1000), GAPDH (PT #60004–1-Ig, 1:4000).

Techniques: Expressing, Western Blot

Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ mitigates mitochondrial H 2 O 2 -mediated damage and supports quality control in cardiomyocytes under hypoxia-reoxygenation stress

doi: 10.1016/j.redox.2025.103799

Figure Lengend Snippet: PrxⅢ depletion exacerbates mitochondrial and cytoplasmic H 2 O 2 accumulation under hypoxia/reoxygenation (H/R) stress in cardiomyocytes. (A) Clone validation for PrxⅢ knockdown. H9c2 cardiomyocytes were transfected with either pSUPER (control) or pSUPER-siPrxⅢ vectors, and stable clones were selected by puromycin resistance. Western blotting was performed on six independent clone sets to evaluate PrxⅢ expression, with β-actin used as a loading control. (B) Protein expression of mitochondrial antioxidant enzymes (Trx2, SOD2, and Gpx4) in pSUPER and pSUPER-siPrxⅢ cells under basal conditions. (C) Validation of hypoxia/reoxygenation conditions using HIF-1α protein levels. Cells were subjected to normoxia (Control), hypoxia for 1 h (H), or hypoxia followed by 4 h of reoxygenation (H/R), and lysates were immunoblotted for HIF-1α and β-actin. (D) Mitochondrial H 2 O 2 was measured using MitoPY-1 and quantified by flow cytometry. (E) Cytoplasmic H 2 O 2 was detected using the PO-1 fluorescent probe and analyzed by flow cytometry. Representative histograms (left) and quantification of relative fluorescence intensity (RFI, %) (right) are shown for D and E. All data are expressed as mean ± S.D. from independent biological replicates (n = 5). Statistical significance was determined using two-way ANOVA followed by Bonferroni's post hoc test. ∗p < 0.05 and ∗∗∗p < 0.001.

Article Snippet: The primary antibodies against poly(ADP-ribose) polymerase (PARP) (9542), cleaved caspase-3 (9661s), caspase-9 (9508), pDrp1 (Ser616; 3455s), pDrp1 (Ser637; 6319s), light-chain 3B (LC3B) (2775s), SQSTM1/p62 (5114T), lysosome-associated membrane protein 1 (LAMP1) (9091s), hypoxia-inducible factor-1 α (HIF-1α) (14179s), thioredoxin 2 (Trx2) (14907s) and HSP60 (12156) were purchased from Cell Signaling Technology (CST, Danvers, MA, USA); 8-hydroxy-2′-deoxyguanosine (8-OHdG) (SC66036), Parkin (PRK8l SC32282), Bcl2 adenovirus e1B 19 kDa interacting protein 3 (BNIP3) (SC56167), Drp1 (SC271583), and mitofusin 1 (Mfn1) (SC166644) were purchased from Santa Cruz Biotechnologies (Santa Cruz, Dallas, TX, USA); optic atrophy 1 (OPA1) (ab42364) was purchased from Abcam (Cambridge, UK); cytochrome C (556433) was purchased from BD Pharmingen (San Jose, CA, USA); catalase (LF-PA0060), Gpx4 (LF-PA0055), SOD2 (LF-PA0021), β-actin (LF-PA0207) and PrxIII (mono; LF-MA0043) were purchased from Ab Frontier (Seoul, Republic of Korea); glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (MAB374) was purchased from Chemicon (Jincheon, Republic of Korea); The secondary antibodies against anti-mouse IgG HRP (474–1806) and anti-rabbit IgG HRP (5220-0458) were purchased from Seracare (Milford, MA, USA).

Techniques: Biomarker Discovery, Knockdown, Transfection, Control, Clone Assay, Western Blot, Expressing, Flow Cytometry, Fluorescence