Journal: Materials Today Bio

Article Title: One-step green assembly of natural polyphenol-based nobiletin nanoparticles for ulcerative colitis therapy

doi: 10.1016/j.mtbio.2026.102887

Figure Lengend Snippet: | In vivo anti-oxidant and anti-inflammatory effects of NTAl NPs against UC. A) H&E-stained sections of mouse colon following treatment with indicated materials. B) H&E staining histological scores. C) Typical TEM images showing the microstructures of colonic epitheliums. D-F) the expression of oxidative stress markers including MDA(D), SOD (E), and GSH-Px (F). G-J) the expression of colonic inflammatory cytokines including pro-inflammatory cytokines TNF-α (G), IL-1β (H), IL-6 (I) and anti-inflammatory cytokines IL-10 (J). All data are shown as mean ± s.d; n = 6. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Commercial ELISA kits including Mouse TNF-α (EK282, Multi Sciences), Mouse IL-1β (EK201B, Multi Sciences), Mouse IL-6 (EK206, Multi Sciences), and Mouse IL-10 (EK210, Multi Sciences) were used to quantify pro-inflammatory cytokines and the anti-inflammatory cytokine according to the manufacturer's instructions.

Techniques: In Vivo, Staining, Expressing

Journal: JBMR Plus

Article Title: USP34 attenuates cartilage degradation in temporomandibular joint osteoarthritis by ANT1-mediated mitophagy

doi: 10.1093/jbmrpl/ziag004

Figure Lengend Snippet: Deletion of Usp34 accelerates cartilage destruction during TMJ OA. (A) Representative images of Safranin O/Fast Green staining of SHAM and UBR-induced TMJ OA mice. Scale bars: 100 μm for low and 50 μm for high magnification. (B and C) Quantitative analysis regarding cartilage thickness and modified Mankin score according to Safranin O/Fast Green staining. n = 6 per group. (D) Representative micro-CT images reveal the subchondral bone microstructure. Scale bars: 100 μm. (E) Quantitative analysis of subchondral bone parameters. (F and G) Representative images and quantitative analysis of western blot for MMP13, ADAMTS5, α-tubulin in ATDC5 cells transfected with Usp34 siRNA following exposure to IL-1β. (H) Relative mRNA expression of MMP3, MMP13, ADAMTS4, and ADAMTS5 in ATDC5 cells transfected with Usp34 siRNA following exposure to IL-1β.

Article Snippet: To model the inflammatory microenvironment of TMJ OA, ATDC5 cells (1 × 10 5 /well) were cultured in 24-well plates, followed by treatment with 10 ng/mL IL-1β (HY-P7073A; MedChemExpress).

Techniques: Staining, Modification, Micro-CT, Western Blot, Transfection, Expressing

Journal: JBMR Plus

Article Title: USP34 attenuates cartilage degradation in temporomandibular joint osteoarthritis by ANT1-mediated mitophagy

doi: 10.1093/jbmrpl/ziag004

Figure Lengend Snippet: USP34 deubiquitinates and stabilizes ANT1. (A) Volcano plots showing the differentially expressed proteins of USP34-deficient cells from public proteomic dataset in the National Genomics Data Center under accession numbers: OMIX007639. (B) Heatmap showing the differentially expressed protein of USP34-deficient cells from public proteomic dataset (OMIX007639). (C and D) Representative images and quantitative analysis of western blot for ANT1 and α-tubulin in ATDC5 cells transfected with Usp34 siRNA following exposure to IL-1β. (E) Representative images of immunofluorescence staining of ANT1 in the TMJ cartilages of SHAM and UBR-induced TMJ OA mice. Scale bars: 50 μm. (F) Co-immunoprecipitation of USP34 with ectopically expressed ANT1 in HEK293T cells. (G) Immunoblot of ANT1-linked polyubiquitin. HEK293T cells were treated with 10 μM MG132 for 4 h after transfection with the indicated constructs. The cell lysates were subjected to immunoprecipitation with the indicated antibody. (H) Measurement of ANT1 degradation rate. HEK293T cells were transfected with the indicated constructs and treated with 10 mg/mL CHX.

Article Snippet: To model the inflammatory microenvironment of TMJ OA, ATDC5 cells (1 × 10 5 /well) were cultured in 24-well plates, followed by treatment with 10 ng/mL IL-1β (HY-P7073A; MedChemExpress).

Techniques: Western Blot, Transfection, Immunofluorescence, Staining, Immunoprecipitation, Construct

Journal: JBMR Plus

Article Title: USP34 attenuates cartilage degradation in temporomandibular joint osteoarthritis by ANT1-mediated mitophagy

doi: 10.1093/jbmrpl/ziag004

Figure Lengend Snippet: ANT1 overexpression rescues mitochondrial homeostasis in USP34-deficient cells. (A and B) Representative images and quantitative analysis of western blot for LC3, Parkin, PINK1, and α-tubulin in ATDC5 cells transfected with Usp34 siRNA or Lv-ANT1 , following exposure to IL-1β. (C) Representative TEM images of ATDC5 cells transfected with Usp34 siRNA or Lv-ANT1 . Subcellular structures with discernible mitochondria (yellow arrows) and bound by a double limiting membrane are identified as putative mitophagosome structures (red arrows). Scale bars: 200 nm. (D) ATDC5 cells stained with mitotracker and lysotracker after transfection with Usp34 siRNA or Lv-ANT1 . Scale bars: 20 μm. (E) ATDC5 cells stained with Mito-SOX after transfection with Usp34 siRNA or Lv-ANT1 . Scale bars: 20 μm.

Article Snippet: To model the inflammatory microenvironment of TMJ OA, ATDC5 cells (1 × 10 5 /well) were cultured in 24-well plates, followed by treatment with 10 ng/mL IL-1β (HY-P7073A; MedChemExpress).

Techniques: Over Expression, Western Blot, Transfection, Membrane, Staining

Journal: JBMR Plus

Article Title: USP34 attenuates cartilage degradation in temporomandibular joint osteoarthritis by ANT1-mediated mitophagy

doi: 10.1093/jbmrpl/ziag004

Figure Lengend Snippet: USP34 overexpression enhanced chondrocyte viability. (A and B) Representative images and quantitative analysis of western blot for LC3, Parkin, PINK1, and α-tubulin in ATDC5 cells transfected with Usp34 lentiviral activation particles ( Usp34 ac) following exposure to IL-1β. (C and D) Representative images and quantitative analysis of western blot for MMP13, ADAMTS5, and α-tubulin in ATDC5 cells transfected with Usp34 ac following exposure to IL-1β. (E and F) Representative images and quantitative analysis of western blot for MMP13, ADAMTS5, and α-tubulin in ATDC5 cells with the indicated treatment. (G) ATDC5 cells stained with Acan and Col2a1 after transfection with Usp34 ac or Lv-ANT1 . Scale bars: 50 μm. (H and I) Relative mRNA expression of Acan and Col2a1 in ATDC5 cells with the indicated treatments.

Article Snippet: To model the inflammatory microenvironment of TMJ OA, ATDC5 cells (1 × 10 5 /well) were cultured in 24-well plates, followed by treatment with 10 ng/mL IL-1β (HY-P7073A; MedChemExpress).

Techniques: Over Expression, Western Blot, Transfection, Activation Assay, Staining, Expressing

Journal: International Journal of Cardiology. Cardiovascular Risk and Prevention

Article Title: The sphingolipid metabolite sphingosine protects against hypertension by targeting metabolic-inflammatory crosstalk via the NLRP3 inflammasome

doi: 10.1016/j.ijcrp.2025.200562

Figure Lengend Snippet: SPH attenuates pathological changes and suppresses NLRP3 inflammasome activation in the hearts of hypertensive mice. (A, B) Masson's trichrome staining of heart (A) and aorta (B) sections, showing reduced collagen deposition (blue) with SPH treatment. Scale bars: 20 μm for heart, 50 μm (main) and 20 μm (inset) for aorta. (C) H&E staining of heart sections showing amelioration of myocardial disarray and inflammation. Scale bars: 500 μm (main) and 20 μm (inset).(D) ELISA quantification of serum IL-18 and IL-1β levels. Data are mean ± SEM. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001 vs. HBP group. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Serum levels of interleukin-18 (IL-18) and interleukin-1β (IL-1β) were quantified using commercial enzyme-linked immunosorbent assay (ELISA) kits (IL-18: Lianke, Cat#EK218-96; IL-1β: Lianke, Cat# EK201BHS-96) according to the manufacturers' instructions.

Techniques: Activation Assay, Staining, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Cardiology. Cardiovascular Risk and Prevention

Article Title: The sphingolipid metabolite sphingosine protects against hypertension by targeting metabolic-inflammatory crosstalk via the NLRP3 inflammasome

doi: 10.1016/j.ijcrp.2025.200562

Figure Lengend Snippet: SPH mitigates Angiotensin II-induced inflammasome activation, apoptosis, and oxidative stress in HUVECs. (A, B) Representative immunofluorescence images showing increased expression of NLRP3 (red, A) and ASC (green, B) in AngII-treated cells, which is reduced by subsequent SPH treatment. Scale bar = 100 μm. (C) TUNEL assay (green) demonstrating increased apoptosis in AngII-treated cells, which is attenuated by SPH. Scale bar = 100 μm. (D, E) Biochemical assays showing that SPH or MCC950 treatment reverses the AngII-induced decrease in SOD1 activity (G).NO (H) and increase in MDA levels (D). (F, G) DCFH-DA staining showing increased intracellular ROS (green) after AngII treatment, which is suppressed by SPH or MCC950. Representative images (F) and quantification (E) are shown. (I, J) ELISA results showing that SPH or MCC950 treatment inhibits the AngII-induced secretion of IL-1β (I) and IL-18 (J). (K) Scanning electron microscopy (SEM) images showing that SPH or MCC950 treatment improves the cell surface morphology and reduces features of pyroptotic damage induced by AngII. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Serum levels of interleukin-18 (IL-18) and interleukin-1β (IL-1β) were quantified using commercial enzyme-linked immunosorbent assay (ELISA) kits (IL-18: Lianke, Cat#EK218-96; IL-1β: Lianke, Cat# EK201BHS-96) according to the manufacturers' instructions.

Techniques: Activation Assay, Immunofluorescence, Expressing, TUNEL Assay, Activity Assay, Staining, Enzyme-linked Immunosorbent Assay, Electron Microscopy

Journal: PLOS One

Article Title: A new mechanism regulating microglial NLRP3 inflammasome: FMR1 mediates NLRP3 mRNA stability

doi: 10.1371/journal.pone.0341867

Figure Lengend Snippet: (A) qPCR analysis of NLRP3 mRNA in BV2 cells treated with vehicle (PBS), LPS/ATP, or LPS/ATP + MCC950. Data are expressed as mean ± SD of 3 biologically independent experiments. (B) Representative Western blots and quantification of NLRP3, pro-caspase-1, and cleaved caspase-1 (p20) protein levels. β-actin served as a loading control. Data are expressed as mean ± SD of 3 biologically independent experiments. (C) Caspase-1 activity measured using a commercial assay kit. Data are expressed as mean ± SD of 3 biologically independent experiments. (D and E) Secretion levels of mature IL-1β and IL-18 in cell culture supernatants determined by ELISA. Data are expressed as mean ± SD of 3 biologically independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s. non-significant.

Article Snippet: The levels of IL-1β and IL-18 in the supernatant of treated BV2 microglia were determined using Mouse IL-1β ELISA Kit (#EK201B) and Mouse IL-18 ELISA Kit (#EK218), respectively, as described by the vendor (Multi Sciences, Hangzhou, China).

Techniques: Western Blot, Control, Activity Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: PLOS One

Article Title: A new mechanism regulating microglial NLRP3 inflammasome: FMR1 mediates NLRP3 mRNA stability

doi: 10.1371/journal.pone.0341867

Figure Lengend Snippet: (A) Measurement of NLRP3, pro-caspase-1 and cleaved caspase-1 levels by western blot in BV2 cells transfected with FMR1 expression construct, FMR1 expression construct+NLRP3 cDNA plasmid, or vector control before LPS/ATP exposure. Data are expressed as mean ± SD of 3 biologically independent experiments. (B) Evaluation of caspase-1 activity in BV2 cells treated as indicated using the assay kit. Data are expressed as mean ± SD of 3 biologically independent experiments. (C and D) Determination of IL-1β and IL-18 secretion levels by ELISA in the supernatant of BV2 cells treated as in A. Data are expressed as mean ± SD of 3 biologically independent experiments. (E) BV2 cells were transfected and treated as indicated, followed by LPS/ATP stimulation. The release of LDH into the culture supernatant was quantified as a marker of cell membrane integrity loss during pyroptosis. Data are expressed as mean ± SD of 3 biologically independent experiments. (F) FMR1 attenuates LPS/ATP-induced pyroptosis, as assessed by Calcein-AM/PI staining. Representative fluorescent micrographs (left) and quantification data (right). Nuclei were counterstained with DAPI (blue). Scale bar, 100 μm. Data are expressed as mean ± SD of 3 biologically independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The levels of IL-1β and IL-18 in the supernatant of treated BV2 microglia were determined using Mouse IL-1β ELISA Kit (#EK201B) and Mouse IL-18 ELISA Kit (#EK218), respectively, as described by the vendor (Multi Sciences, Hangzhou, China).

Techniques: Western Blot, Transfection, Expressing, Construct, Plasmid Preparation, Control, Activity Assay, Enzyme-linked Immunosorbent Assay, Marker, Membrane, Staining

Journal: iScience

Article Title: Maimendong decoction suppresses non-small cell lung cancer growth by promoting dendritic cell maturation via the SIRT1/p65 acetylation pathway

doi: 10.1016/j.isci.2026.114774

Figure Lengend Snippet: MMDD suppresses tumor growth via SIRT1 downregulation and enhanced p65 acetylation (A and C) Western blot analysis of SIRT1 and acetyl-p65 protein levels in tumor tissues from vehicle control (sterile deionized water), L-MMDD, M-MMDD, and H-MMDD groups. GAPDH served as the internal loading control for SIRT1, acetyl-p65, and p65. (B and D) Quantitative analysis of relative SIRT1 and acetyl-p65/p65 expression levels in tumor tissues using ImageJ software ( n = 3). (E) Representative immunofluorescence images of SIRT1 (red) and acetyl-p65 (green) in tumor sections. Nuclei were counterstained with DAPI (blue) (scale bar, 50 μm). (F) Quantification of mean fluorescence intensity for SIRT1 and acetyl-p65 ( n = 3). (G) ELISA quantification of cytokine levels (IL-1β, IL-6, IL-12, TNF-α, and IFN-γ) in tumor homogenates ( n = 6). Data are represented as mean ± SD. One-way ANOVA was used for comparison among multiple groups.∗ p < 0.05 and ∗∗ p < 0.01.

Article Snippet: ELISA Kits (IL-1β, IL-6, IL-12, TNF-α, IFN-γ) , LiankeBio , Cat# EK201B; EK206; EK2183; EK282; EK280.

Techniques: Western Blot, Control, Sterility, Expressing, Software, Immunofluorescence, Fluorescence, Enzyme-linked Immunosorbent Assay, Comparison

Journal: iScience

Article Title: Maimendong decoction suppresses non-small cell lung cancer growth by promoting dendritic cell maturation via the SIRT1/p65 acetylation pathway

doi: 10.1016/j.isci.2026.114774

Figure Lengend Snippet: MMDD-containing serum-treated DCs enhance the migration, invasion, and activation of CD8 + T cells (A) Representative transwell assay images show the migration and invasion capacities of CD8 + T cells co-cultured with DCs pretreated with control (drug-free serum from vehicle-treated rats), low-, medium-, or high-dose MMDD-containing serum (scale bar, 50 μm). (B) Quantification of migrated and invaded CD8 + T cells ( n = 3). (C) Flow cytometry analysis of IFN-γ expression in CD8 + T cells co-cultured with supernatant of MMDD-treated DCs. (D) Quantification of the proportion of IFN-γ + cells among CD8 + T cells ( n = 3). Data are represented as mean ± SD. One-way ANOVA (D) and two-way ANOVA (B) are used for comparison among multiple groups.∗ p < 0.05 and ∗∗ p < 0.01.

Article Snippet: ELISA Kits (IL-1β, IL-6, IL-12, TNF-α, IFN-γ) , LiankeBio , Cat# EK201B; EK206; EK2183; EK282; EK280.

Techniques: Migration, Activation Assay, Transwell Assay, Cell Culture, Control, Flow Cytometry, Expressing, Comparison

Journal: iScience

Article Title: Maimendong decoction suppresses non-small cell lung cancer growth by promoting dendritic cell maturation via the SIRT1/p65 acetylation pathway

doi: 10.1016/j.isci.2026.114774

Figure Lengend Snippet: MMDD exerts antitumor activity by modulating the SIRT1/acetyl-p65 axis in DCs (A and C) Western blot analysis of SIRT1, acetyl-p65, and p65 expression in DCs treated with control (drug-free serum from vehicle-treated rats), low-, medium-, or high-dose MMDD-containing serum. GAPDH served as the internal loading control for SIRT1, acetyl-p65, and p65. (B and D) Quantitative analysis of relative SIRT1 and acetyl-p65/p65 expression levels in DCs using ImageJ software ( n = 3). (E) ELISA quantification of pro-inflammatory cytokines (IL-1β, IL-6, IL-12, and TNF-α) and IFN-γ in supernatants of MMDD-treated DCs ( n = 6). (F) Western blot analysis of SIRT1 expression in control, negative control (NC), and AAV-SIRT1-transduced (OE-SIRT1) BMDCs. (G) Quantification of SIRT1 expression levels in DCs ( n = 3). (H) Western blot analysis of SIRT1, acetyl-p65, and p65 in DCs from the control, high-dose MMDD-containing serum, OE-SIRT1, or OE-SIRT1+high-dose MMDD-containing serum groups. (I and J) Quantification of relative SIRT1, acetyl-p65/p65 levels in DCs ( n = 3). (K) ELISA analysis of pro-inflammatory cytokines (IL-1β, IL-6, IL-12, and TNF-α) and IFN-γ in DC supernatants ( n = 6). (L) Flow cytometry profiles of DCs maturation markers (CD40, CD80, CD83, and MHC-II) across groups. (M) Quantitative analysis of CD40 + , CD80 + , CD83 + , and MHC-II + DC proportions ( n = 3). (N) Flow cytometry analysis of IFN-γ expression in CD8 + T cells treated with DC-conditioned supernatant from different groups. (O) Quantification of the proportion of IFN-γ + cells among CD8 + T cells ( n = 3). Data are represented as mean ± SD. One-way ANOVA was used for comparison among multiple groups.∗ p < 0.05 and ∗∗ p < 0.01. See also .

Article Snippet: ELISA Kits (IL-1β, IL-6, IL-12, TNF-α, IFN-γ) , LiankeBio , Cat# EK201B; EK206; EK2183; EK282; EK280.

Techniques: Activity Assay, Western Blot, Expressing, Control, Software, Enzyme-linked Immunosorbent Assay, Negative Control, Flow Cytometry, Comparison