Journal: Frontiers in Immunology

Article Title: Downregulation of Cathepsin B expression alleviates periodontitis by reducing mitochondrial reactive oxygen species production and NOD-, LRR-, and pyrin domain-containing 3 -mediated pyroptosis

doi: 10.3389/fimmu.2026.1762290

Figure Lengend Snippet: AAV-CTSB alleviates alveolar bone destruction in mice periodontitis. (A) Three-dimensional reconstruction images and sagittal plane images of the maxillary bone categorized by group. The red shaded area represents the amount of alveolar bone resorption, and the white dotted line represents the distance from the cementoenamel junction (CEJ) to the alveolar bone crest (ABC). Quantitative analysis of (B) the bone resorption area and (C) distance from CEJ to ABC. NS, not significant, ** p <.01, *** p <.001.

Article Snippet: Subsequently, AAV-CTSB (AAV‐U6‐CTSB‐shRNA‐CAG‐EGFP; GeneChem) was transfected in the mice periodontal region for 21 days to downregulate CTSB expression ( ).

Techniques:

Journal: Frontiers in Immunology

Article Title: Downregulation of Cathepsin B expression alleviates periodontitis by reducing mitochondrial reactive oxygen species production and NOD-, LRR-, and pyrin domain-containing 3 -mediated pyroptosis

doi: 10.3389/fimmu.2026.1762290

Figure Lengend Snippet: AAV-CTSB reduces macrophage infiltration in periodontal lesions. (A) Immunohistochemistry (IHC) analysis of CTSB in the periodontal area of each group. (B) IHC staining of F4/80 in the periodontal area of each group. (C) Quantitative analysis of CTSB-positive cells. (D) Quantification of F4/80-positive cells in each group. NS, not significant, ** p <.01, *** p <.001.

Article Snippet: Subsequently, AAV-CTSB (AAV‐U6‐CTSB‐shRNA‐CAG‐EGFP; GeneChem) was transfected in the mice periodontal region for 21 days to downregulate CTSB expression ( ).

Techniques: Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: Downregulation of Cathepsin B expression alleviates periodontitis by reducing mitochondrial reactive oxygen species production and NOD-, LRR-, and pyrin domain-containing 3 -mediated pyroptosis

doi: 10.3389/fimmu.2026.1762290

Figure Lengend Snippet: AAV-CTSB inhibits NLRP3-mediated pyroptosis and the production of inflammatory factors in mice periodontitis. (A) Immunofluorescence (IF) staining of CTSB in the periodontal tissues of each group. (B) Quantification of CTSB-positive cells in each group. (C) Relative mRNA expression levels of Ctsb, Nlrp3, Casp1, Tnfα, and Il1β in the periodontal tissues of each group as detected by quantitative real-time polymerase chain reaction. (D) Interleukin-1β concentration as detected by enzyme-linked immunosorbent assay. NS, not significant, *p<.05, **p<.01, ***p<.001. .

Article Snippet: Subsequently, AAV-CTSB (AAV‐U6‐CTSB‐shRNA‐CAG‐EGFP; GeneChem) was transfected in the mice periodontal region for 21 days to downregulate CTSB expression ( ).

Techniques: Immunofluorescence, Staining, Expressing, Real-time Polymerase Chain Reaction, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Downregulation of Cathepsin B expression alleviates periodontitis by reducing mitochondrial reactive oxygen species production and NOD-, LRR-, and pyrin domain-containing 3 -mediated pyroptosis

doi: 10.3389/fimmu.2026.1762290

Figure Lengend Snippet: AAV-CTSB decreases NLRP3-mediated pyroptosis in mice macrophages. (A) Immunofluorescence staining of F4/80 and NLRP3 in the periodontal tissues of each group. (B) Quantification of F4/80-positive cells in each group. (C) Quantitative analysis of NLRP3-positive cells. NS, not significant, * p <.05, ** p <.01, *** p <.001.

Article Snippet: Subsequently, AAV-CTSB (AAV‐U6‐CTSB‐shRNA‐CAG‐EGFP; GeneChem) was transfected in the mice periodontal region for 21 days to downregulate CTSB expression ( ).

Techniques: Immunofluorescence, Staining

Journal: Frontiers in Immunology

Article Title: Downregulation of Cathepsin B expression alleviates periodontitis by reducing mitochondrial reactive oxygen species production and NOD-, LRR-, and pyrin domain-containing 3 -mediated pyroptosis

doi: 10.3389/fimmu.2026.1762290

Figure Lengend Snippet: AAV-CTSB alleviates NLRP3-mediated pyroptosis in in vitro experiments. (A, B) western blotting and quantification analysis of CTSB, NLRP3, pro-caspase-1, cleaved-caspase-1, gasdermin-D (GSDMD), and GSDMD-N expression in the macrophage cell lines of each group. (C) Relative mRNA expression levels of Ctsb, Nlrp3, Casp1, and Gsdmd in macrophages as detected by quantitative real-time polymerase chain reaction. NS, not significant, * p <.05, ** p <.01, *** p <.001.

Article Snippet: Subsequently, AAV-CTSB (AAV‐U6‐CTSB‐shRNA‐CAG‐EGFP; GeneChem) was transfected in the mice periodontal region for 21 days to downregulate CTSB expression ( ).

Techniques: In Vitro, Western Blot, Expressing, Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: Downregulation of Cathepsin B expression alleviates periodontitis by reducing mitochondrial reactive oxygen species production and NOD-, LRR-, and pyrin domain-containing 3 -mediated pyroptosis

doi: 10.3389/fimmu.2026.1762290

Figure Lengend Snippet: AAV-CTSB reduces mitochondrial reactive oxygen species (ROS) production and pyroptosis downstream inflammatory factors. (A) MitoSOX and 2’-7’-dichlorodihydrofluorescein diacetate (DCFH-DA) staining applied to detect mitochondrial and intracellular ROS production. (B) Quantitative analysis of relative MitoSOX red fluorescence intensity in each group. (C) Quantitative analysis of relative DCFH-DA fluorescence intensity in each group. (D) Relative mRNA expression levels of Il18, Il1β, Tnfα, and Il6 in macrophages as detected by quantitative real-time polymerase chain reaction. (E) The expression level of interleukin-1β in the cell supernatant of different groups as detected by enzyme-linked immunosorbent assay. NS, not significant, ** p <.01, *** p <.001.

Article Snippet: Subsequently, AAV-CTSB (AAV‐U6‐CTSB‐shRNA‐CAG‐EGFP; GeneChem) was transfected in the mice periodontal region for 21 days to downregulate CTSB expression ( ).

Techniques: Staining, Fluorescence, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Journal of Advanced Research

Article Title: Integrative multi-omics profiling deciphers tumor microenvironment heterogeneity and immunotherapy vulnerabilities in lung neuroendocrine carcinomas

doi: 10.1016/j.jare.2025.06.017

Figure Lengend Snippet: Construction and performance evaluation of the iPROM. (A) Schematic overview for developing the Immuno-Proteomic XGBoost model (iPROM). (B) AUC-ROC for the predicted performance between IPC1 and IPC2 in the training set (n = 51) and test set (n = 25). (C) Distribution of the iPROM scores with a threshold of 0.5. Red indicates IPC1-like subtype and grey indicates IPC2-like subtype. (D) AUC-ROC analysis showing the performance of the iPROM in the George cohort (n = 147). Distribution of the iPROM scores with a threshold of 0.5. Red indicates IPC1-like subtype and grey indicates IPC2-like subtype. AUC-PR analysis and bar plots showing the distribution of accuracy, sensitivity, specificity, precision, recall and F1-score in the George cohort. (E-F) Heatmap showing the distribution of CTSB , FCGR2C and FCGR3A between iPROM hi and iPROM lo subtypes in CHCAMS cohort 2 (n = 112) and Roper cohort (n = 17). Kaplan-Meier curves showing the differences in prognosis (OS and DFS) between iPROM subtypes (log-rank test). Bar plots showing the distribution of response status between iPROM hi and iPROM lo subtypes. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The primary antibodies included FCGR2C (HPA010718, Merck; 1:200), FCGR3A (16559-1-AP, Proteintech; 1:100), and CTSB (12216-1-AP, Proteintech; 1:100).

Techniques:

Journal: Biology Direct

Article Title: Obesity-associated lysosomal nitrosative stress promotes retinopathy

doi: 10.1186/s13062-026-00743-1

Figure Lengend Snippet: Obesity-induced nitrosative stress impairs lysosomal processing of VEGF in the retina. ( A ) Retinal Vegfa mRNA expression extracted from publicly available GEO datasets. ( B ) VEGF protein levels in retinas from WT and iNOS knockout (KO) mice fed RD or HFD for 24 wks, measured by ELISA and normalized to RD-fed WT controls ( n = 7 mice/group). ( C ) ctsb mRNA expression in retinas from lean and obese mice based on public GEO datasets. ( D – E ) CTSB protein levels ( D ) and enzymatic activity ( E ) in retinas from WT and iNOS KO mice fed RD or HFD, measured by ELISA and enzymatic assay, respectively ( n = 7 mice/group). ( F – G ) Representative immunofluorescence images ( F ) and co-localization analysis ( G ) of S-nitrosylated CTSB in retinal sections from RD- and HFD-fed WT mice. ROS: rod outer segment; RIS: rod inner segment; ONL: outer nuclear layer; OPL: outer plexiform layer; INL: inner nuclear layer; IPL: inner plexiform layer; GCL: ganglion cell layer. -AS: Ascorbat omitted. Scale bars, 20 μm ( H – I ) S-nitrosylation of CTSB assessed by biotin switch assay in retinal lysates ( H ), with densitometric quantification shown in ( I ) ( n = 3 mice/group). Data are presented as mean ± SEM. * p < 0.05 indicates dietary effect within the same genotype; # p < 0.05 indicates genotype effect under the same diet. Statistical tests: Student’s t -test for (A, C, G); one-way ANOVA with post hoc test for (B, D, E, I)

Article Snippet: The wells were blocked in 5% (w/v) bovine serum albumin (BSA) and incubated at room temperature for 2 h. The wells were incubated overnight with an anti-rabbit VEGF polyclonal antibody (1:3000, ThermoFisher), anti-rabbit GSNOR polyclonal antibody (1:5000, Abcam), or anti-rabbit CTSB monoclonal antibody (1:3000, Cell Signaling Technology), anti-rabbit F4/80 monoclonal antibody (1:3000, Cell Signaling Technology) antibodies, followed by washing and then incubation with anti-rabbit IgG, HRP-linked secondary antibody and TMB substrate (BioLegend).

Techniques: Expressing, Knock-Out, Enzyme-linked Immunosorbent Assay, Activity Assay, Enzymatic Assay, Immunofluorescence, Biotin Switch Assay

Journal: Biology Direct

Article Title: Obesity-associated lysosomal nitrosative stress promotes retinopathy

doi: 10.1186/s13062-026-00743-1

Figure Lengend Snippet: GSNOR protects against obesity-associated retinal lysosomal dysfunction. ( A ) GSNOR enzymatic activity in retinas of GSNOR knockout (KO) and WT mice fed an RD ( n = 6 mice/group). ( B – C ) Retinal VEGF ( B ) and CTSB protein levels ( C ) in GSNOR KO and WT mice, assessed by indirect ELISA ( n = 6 mice/group). ( D ) CTSB enzymatic activity in retinas from GSNOR KO and WT mice ( n = 6 mice/group). ( E ) Representative Western blot showing S-nitrosylation of CTSB in retinas from GSNOR KO and WT mice using the biotin switch assay. ( F ) Densitometric analysis of blot data in ( E ). All data are presented as mean ± SEM. * p < 0.05 indicates statistical significance as determined by Student’s t -test

Article Snippet: The wells were blocked in 5% (w/v) bovine serum albumin (BSA) and incubated at room temperature for 2 h. The wells were incubated overnight with an anti-rabbit VEGF polyclonal antibody (1:3000, ThermoFisher), anti-rabbit GSNOR polyclonal antibody (1:5000, Abcam), or anti-rabbit CTSB monoclonal antibody (1:3000, Cell Signaling Technology), anti-rabbit F4/80 monoclonal antibody (1:3000, Cell Signaling Technology) antibodies, followed by washing and then incubation with anti-rabbit IgG, HRP-linked secondary antibody and TMB substrate (BioLegend).

Techniques: Activity Assay, Knock-Out, Indirect ELISA, Western Blot, Biotin Switch Assay