reca 1  (Proteintech)

Journal: Military Medical Research

Article Title: USP18 exacerbates myocardial I/R injury by inhibiting Parkin mitophagy through the deubiquitinase PTEN-L

doi: 10.1016/j.mmr.2026.100004

Figure Lengend Snippet: USP18 aggravates cardiac I/R injury through regulation of mitochondria and inhibition of mitophagy. a Electron microscopy image showing mitophagy in USP18-cKO mouse hearts ( n= 5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. b Protein levels of PINK1, Parkin, ubiquitinated proteins (Ub), P62, and LC3II in mitochondria from heart tissue in USP18-cKO and WT mice 24 h after I/R injury ( n =4). c Electron microscopy image showing mitophagy in USP18-overexpres (OV) mouse hearts ( n =5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. d Protein levels of PINK1, Parkin, Ub, P62, and LC3II proteins in mitochondria from the heart tissue of USP18-OV mice 24 h after I/R injury ( n =4). Color shift in mitophagy dye (red) and lysosomal dye (green) in NRVMs showing mitophagy in NRVMs with USP18 siRNA transfection ( e ) or Ad-USP18 infection ( f ) and the quantitative mitophagy index in each group ( n= 5). Scale bar=9 μm. Protein levels of PINK1, Parkin, Ub, P62, and LC3II in mitochondria from NRVMs transfected with USP18 siRNA ( g ) or infected with Ad-USP18 ( h ). ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001 ⁎⁎⁎⁎ P <0.0001. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; WT. Wild-type; KO. Knockout; P62. Sequestosome 1; LC3. Microtubule-associated protein 1 light chain 3; VDAC. Voltage-dependent anion channel.

Article Snippet: To block mitophagy, the selective dynamin-related protein 1 (Drp1) inhibitor Mdivi-1 was used (50 μmol/L, MedChemExpress, USA).

Techniques: Inhibition, Electron Microscopy, Transfection, Infection, Ubiquitin Proteomics, Knock-Out

Journal: Journal of Virology

Article Title: Nectin4 restricts pseudorabies virus infection by blocking gD-Nectin1 interaction

doi: 10.1128/jvi.00788-26

Figure Lengend Snippet: Nectin4 competitively attenuates gD and Nectin1 interaction. ( A ) HEK293T cells were cotransfected with Flag-tagged Nectin1, Myc-empty vector or Myc-PRV-gD, and either full-length or mutant Nectin4 plasmid. Protein samples were lysed at 4°C and incubated with Myc-magnetic beads for 2 h. The Nectin1-PRV-gD interaction was assessed by Western blot. The interaction between Nectin1 and gD was evaluated under different doses of Nectin4 and its mutants. HEK293T cells were cotransfected with Flag-tagged Nectin1, Myc-tagged PRV-gD, and increasing amounts of full-length Nectin4 ( B ), V5-tagged IgV-deletion ( C ), or V5-tagged IgC2 deletion mutants ( D ). The RFP control vector was included as a negative control. Cell lysates were incubated with Myc-magnetic beads at 4°C for 2 h, and the Nectin1-gD interaction was analyzed by Western blot. ( E ) The Nectin1-gD interaction under increasing doses of Nectin4 was determined by SPR. The Nectin4 protein was injected at increasing concentrations onto a surface precoated with a saturated mixture of Nectin1 and PRV-gD proteins. The binding affinity between PRV-gD and Nectin1 was measured by monitoring changes in resonance units (RU) in response to Nectin4 binding.

Article Snippet: The purified human Nectin1 protein was purchased from MedChemExpress (HY- P70494 ).

Techniques: Plasmid Preparation, Mutagenesis, Incubation, Magnetic Beads, Western Blot, Control, Negative Control, Injection, Binding Assay

Journal: Journal of Virology

Article Title: Nectin4 restricts pseudorabies virus infection by blocking gD-Nectin1 interaction

doi: 10.1128/jvi.00788-26

Figure Lengend Snippet: Schematic illustrating the mechanism by which Nectin4 restricts pseudorabies virus infection through blocking gD-Nectin1 interaction. PRV infection involves multiple steps, including attachment, internalization, membrane fusion, and genome release. Initial attachment is primarily mediated through interactions of viral glycoprotein gC with cell-surface heparan sulfate proteoglycans. Receptor binding by gD initiates the fusion cascade and viral internalization. Upon PRV infection, the interferon signaling pathway is activated (left), subsequently upregulating the expression of interferon-stimulated gene Nectin4 in neighboring cells. Nectin4 is located on the cell membrane and binds to gD through its IgV domain, preventing viral internalization (right). Figure created with BioRender (biorender.com).

Article Snippet: The purified human Nectin1 protein was purchased from MedChemExpress (HY- P70494 ).

Techniques: Virus, Infection, Blocking Assay, Membrane, Binding Assay, Expressing

Journal: Bioactive Materials

Article Title: Targeting VEGFR2 inhibition within a spatially-confined conduit promotes nerve self-resolution and alleviates mechanical allodynia

doi: 10.1016/j.bioactmat.2026.03.009

Figure Lengend Snippet: Prevention of neuroma formation by a spatially confined conduit filled with GelMA MAVP MPS. ( A ) Illustration of 3D-printed GelMA MPs loaded with MAVP and the proposed mechanism of action within the neural conduit. ( B ) Representative images and ( C ) quantitative scores of autotomy behavior over 12 weeks (n = 6). ( D ) Representative gait footprints at 12 weeks post-surgery. ( E ) Quantification of left hindlimb stance duration (n = 6) and ( F ) maximum contact area (n = 6). ( G ) IF staining of p-VEGFR2 activation and ( H ) IF staining of neovascularization marker RECA-1. (I) Quantification of p-VEGFR2-positive area percentage (n = 6). and ( J ) quantification of RECA-1-positive area percentage (n = 6). ( K ) Regenerated nerve length measurements (n = 6). Mean values are shown and error bars represent ± s.d., as analyzed by one-way ANOVA followed by the Tukey-Kramer test in ( C , E , F , I , J and K ). Biological replicates were used for all experiments. ns, p > 0.05, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: The following primary antibodies were used for the subsequent steps: anti-Yap (mouse, 1:200, Santa sc-376830); anti-p-VEGFR2 (rabbit, 1:100 Invitrogen, PA5-105765); α-SMA (rabbit, 1:200, Proteintech 14395-1-AP); Reca-1 (mouse, 1:200, Santa sc-52665); anti-CD31 (mouse, 1:200, Santa sc-13537); anti-Ki67 (rabbit, 1:150, Cell Signaling 9129S); anti-NF-200 (mouse, 1:200, Sigma, SAB4200747); anti-MBP (rabbit, 1:200, Abcam ab218011); anti-F4/80 (mouse, 1:200, Santa sc-377009); Iba-1 (rabbit, 1:150, Abcam ab178846); anti-CGRP (rabbit, 1:400, Abcam ab283568); anti-TRPA1 (mouse, 1:200, Santa sc-376495); anti-CD86 (rabbit, 1:200, Proteintech 30691-1-AP); CD206 (rabbit, 1:200, Proteintech 18704-1-AP).

Techniques: Staining, Activation Assay, Marker

Journal: Marine Drugs

Article Title: Fucoidan Attenuates Lead-Induced Liver Injury Associated with IGFBP1 and Gut Microbiota-Derived Tryptophol Metabolism

doi: 10.3390/md24070232

Figure Lengend Snippet: Integrated transcriptomic and network toxicology analyses identify potential molecular targets of FU against Pb-induced liver injury. ( A ) MA plots showing differentially expressed genes in the comparisons of Model versus Control and FU-H versus Model; ( B ) Venn diagram showing the overlap of differentially expressed genes between the Model versus Control and FU-H versus Model comparisons. A total of 111 common differentially expressed genes were identified; ( C ) KEGG pathway enrichment analysis of the overlapping differentially expressed genes. Bubble size represents the number of enriched genes, and bubble color represents the adjusted p value; ( D ) Venn diagram showing the intersection between Pb-related targets and liver injury-associated targets identified by network toxicology analysis; ( E ) GO enrichment circular plot of the intersecting targets, including biological process, cellular component, and molecular function categories; ( F ) GO Biological Process network illustrating the relationships between enriched biological processes and their associated genes; ( G ) KEGG pathway enrichment analysis of the intersecting targets obtained from network toxicology analysis; ( H ) heatmap showing the expression profiles of MAPK signaling pathway-related differentially expressed genes among the Control, Model, and FU-H groups. IGFBP1 was identified as the most prominently altered candidate gene associated with FU-mediated protection. Differentially expressed genes were screened using the criteria of |log 2 fold change| ≥ 1 and adjusted p < 0.05. Enrichment analyses were performed based on significantly enriched GO terms and KEGG pathways.

Article Snippet: Sodium carboxymethyl cellulose (CMC-Na, HY-Y1889A), recombinant mouse IGFBP1 protein (HY-P700256), and tryptophol (TOL, HY-W010155) were also purchased from MedChemExpress (MCE; Monmouth Junction, NJ, USA).

Techniques: Control, Expressing

Journal: Marine Drugs

Article Title: Fucoidan Attenuates Lead-Induced Liver Injury Associated with IGFBP1 and Gut Microbiota-Derived Tryptophol Metabolism

doi: 10.3390/md24070232

Figure Lengend Snippet: Experimental validation of potential molecular targets involved in FU-mediated protection against Pb-induced liver injury. ( A ) RT-qPCR validation of representative intersecting genes identified by integrated transcriptomic and network toxicology analyses, including Igfbp1 , Ppp1r10 , Dnajb9 , Hspa1a , Cyp2a4 , Cbx2 , Gadd45a , and Egr1 , in the Control, Model, and FU-H groups; ( B – E ) RT-qPCR analysis of MAPK signaling pathway-related genes, including Map2k1 ( B ), Map2k2 ( C ), Mapk1 ( D ), and Mapk3 ( E ). Data are presented as mean ± SD, n = 6 per group. Statistical significance was determined by one-way ANOVA. Compared with the Model group: *** p < 0.001, and **** p < 0.0001.

Article Snippet: Sodium carboxymethyl cellulose (CMC-Na, HY-Y1889A), recombinant mouse IGFBP1 protein (HY-P700256), and tryptophol (TOL, HY-W010155) were also purchased from MedChemExpress (MCE; Monmouth Junction, NJ, USA).

Techniques: Biomarker Discovery, Quantitative RT-PCR, Control

Journal: Marine Drugs

Article Title: Fucoidan Attenuates Lead-Induced Liver Injury Associated with IGFBP1 and Gut Microbiota-Derived Tryptophol Metabolism

doi: 10.3390/md24070232

Figure Lengend Snippet: IGFBP1 is associated with FU-mediated protection against Pb-induced liver injury. ( A ) Schematic illustration of the experimental design for IGFBP1 rescue experiments; ( B ) serum IGFBP1 levels in the Model, FU-H, and IGFBP1+FU-H groups; ( C ) body weight of mice at the end of the experiment; ( D ) liver organ weight index; ( E ) representative H&E-stained liver sections from the Model, FU-H, and IGFBP1+FU-H groups. The upper panels show 100×-magnification images (scale bar = 200 μm), and the lower panels show enlarged views of the dashed areas (scale bar = 100 μm). Red arrows indicate inflammatory cell infiltration, and green arrows indicate fatty vacuolation; ( F ) Histology scores of liver sections. ( G – I ) Serum levels of hepatic injury biomarkers, including ALT ( G ), AST ( H ), and ALP ( I ); ( J – L ) serum oxidative stress-related indicators, including SOD activity ( J ), GSH level ( K ), and MDA level ( L ); ( M – O ) hepatic oxidative stress-related indicators, including SOD activity ( M ), GSH level ( N ), and MDA level ( O ). ( P – R ) Serum levels of pro-inflammatory cytokines, including IL-6 ( P ), TNF-α ( Q ), and IL-1β ( R ); Data are presented as mean ± SD, n = 6 per group. Statistical significance was determined by one-way ANOVA. Compared with the FU-H group: * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Sodium carboxymethyl cellulose (CMC-Na, HY-Y1889A), recombinant mouse IGFBP1 protein (HY-P700256), and tryptophol (TOL, HY-W010155) were also purchased from MedChemExpress (MCE; Monmouth Junction, NJ, USA).

Techniques: Staining, Activity Assay

Journal: Marine Drugs

Article Title: Fucoidan Attenuates Lead-Induced Liver Injury Associated with IGFBP1 and Gut Microbiota-Derived Tryptophol Metabolism

doi: 10.3390/md24070232

Figure Lengend Snippet: Schematic illustration of the potential mechanisms associated with FU-mediated attenuation of Pb-induced liver injury. Pb exposure disrupts gut microbiota homeostasis, induces microbial dysbiosis, and increases systemic Pb burden, thereby contributing to hepatic oxidative stress, inflammatory responses, and liver injury. FU treatment decreased Pb levels in serum and liver and increased fecal Pb content. In the intestine, FU treatment was associated with remodeling of Pb-disrupted gut microbiota, including increased relative abundance of beneficial bacteria such as Akkermansia muciniphila and Bifidobacterium pseudolongum, as well as alterations in fecal tryptophan metabolism, particularly tryptophol (TOL). Through the gut–liver axis, these FU-associated intestinal changes may contribute to the alleviation of Pb-induced hepatic injury. In the liver, FU protection was associated with IGFBP1-related redox modulation, increased antioxidant capacity, including SOD activity and GSH content, reduced lipid peroxidation, as reflected by decreased MDA levels and suppressed inflammatory responses, including TNF-α, IL-1β, and IL-6. These coordinated changes were accompanied by reduced serum ALT, AST, and ALP levels, alleviated hepatic pathological lesions, and improved hepatic homeostasis.

Article Snippet: Sodium carboxymethyl cellulose (CMC-Na, HY-Y1889A), recombinant mouse IGFBP1 protein (HY-P700256), and tryptophol (TOL, HY-W010155) were also purchased from MedChemExpress (MCE; Monmouth Junction, NJ, USA).

Techniques: Bacteria, Activity Assay

Journal: Bioactive Materials

Article Title: Spatiotemporally engineered microneedle for microenvironment remodeling propels mucosal regeneration after tracheal mucosal injury

doi: 10.1016/j.bioactmat.2026.01.026

Figure Lengend Snippet: Gel-AgNA/MgGA MN promote mucosal regeneration. (a) HE and Safranin O staining of rabbit tracheal samples harvested at Day 10 post-operation after treated with Gel, Gel-AgNA, Gel-MgGA, and Gel-AgNA/MgGA MN. (b) IF staining of CK14 (marker of basal cells, red) and AC-Tub (marker of cilia cell, green). (c) IF staining of ZO-1 (marker of tight junctions, orange). (d, e) Quantitative analysis of regenerated epithelial coverage and thickness (n = 9). (f) Masson and Sirius Red staining for collagen evaluation after various treatments (n = 5). Quantitative analysis of collagen volume fraction (g) and fiber orientation (h) . The pentagram indicates luminal side of trachea.

Article Snippet: Immunofluorescence staining of CK14 (Abcam, ab181595), AC-Tub (Proteintech, 66200-1-Ig), ZO-1 (Proteintech, 21773-1-AP), and Immunohistochemical (IHC) staining for CD31 (Servicebio, S1002) were conducted to reveal the conditions of mucosal regeneration, according to previous literature [ ].

Techniques: Staining, Marker