Journal: Bioactive Materials

Article Title: A catalytically active and recyclable bioelastomer inspired by metalloenzymes

doi: 10.1016/j.bioactmat.2026.02.053

Figure Lengend Snippet: Host responses to mouse subcutaneous implantation of Cu-PIAS. ( A ) Representative hematoxylin and eosin staining of the implant site after 7, 14, and 21 days. 25-Cu-PIAS mesh is roughly centered in each image and runs horizontally. Mesh material stains homogenously red and is indicated by arrows in the leftmost image of each timepoint. ( B ) Area of 25-Cu-PIAS mesh at each timepoint. Reduction over time indicates degradation of material. ( C ) Blood vessel density around and within the implant sites (CD31 immunohistochemistry) at each timepoint. ( D ) Total macrophage density around and within the implant sites (F4/80 immunohistochemistry) at each timepoint. ( E ) M2 macrophage density around and within the implant sites (CD163 immunohistochemistry) at each timepoint. ( F ) Ratio of macrophages expressing a M2 phenotype at each timepoint. ( G ) Collagen thickness above the implant site of scarring mouse model (CXCR3 −/− ), comparing responses to 15-Cu-PIAS and fibrin gel after 3 and 6 months. Statistics: ∗, ∗∗, ∗∗∗, and ∗∗∗∗ indicate p ≤ 0.05, 0.01, 0.001, and 0.0001, respectively.

Article Snippet: F4/80 antibody was purchased from Cell Signaling Technology (Danvers MA, Cat #: 70076).

Techniques: Staining, Immunohistochemistry, Expressing

Journal: Journal of Advanced Research

Article Title: Podocyte TLR4 deletion alleviates diabetic kidney disease through prohibiting PKCδ/SHP-1-dependent ER stress and relieving podocyte damage and inflammation

doi: 10.1016/j.jare.2025.07.013

Figure Lengend Snippet: Blockade of TLR4/MyD88/PKCδ/SHP-1 signaling attenuated podocyte cytoskeletal remodeling and inflammation, and macrophage recruitment and infiltration through decreasing ER stress and MCP-1 production under hyperglycemic conditions. (A) Assessment of podocyte migration by transwell assay in cultured podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, MCP-1 or scrambled shRNAs, or pretreatment of 4-PBA. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ## P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments). (B) Representative IF images by phalloidin staining to illustrate F-actin distribution and cytoskeletal remodeling in cultured podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, MCP-1 or scrambled shRNAs, or pretreatment of 4-PBA. Magnification 400 × . ( n = 3 independent experiments). (C) Assessment of the secretion of inflammatory factors IL-6 and IL-1β in podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, or scrambled shRNAs, or pretreatment of 4-PBA. P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.01; ### P < 0.01 ( n = 3 independent experiments). (D) IHC staining using anti-f4/80 antibodies to illustrate macrophage infiltration in the glomeruli from mice at 16 weeks (Red arrow). Scale bar, 20 μm. ( n = 8) (E) The model diaphragm of podocyte-macrophage co-culture. (F) Representative images and assessment of macrophage migration by transwell assay after co-cultured with podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, MCP-1 or scrambled shRNAs, or pretreatment of 4-PBA. Magnification 400 × . P values were determined by one-way ANOVA followed by Tukey’s multiple comparisons test and data are presented as mean ± SD. **** P < 0.01; ## P < 0.01; ### P < 0.01 ( n = 3 independent experiments).

Article Snippet: The following primary antibodies were used: f4/80 (29414–1-AP), desmin (16520–1-AP), MCP-1 (26161–1-AP), podocin (20384–1-AP), synaptopodin (21064–1-AP), TLR4 (66350–1-Ig), β-actin (66009–1-Ig), PKCδ(14188–1-AP), ATF6 (24169–1-AP), p-cadherin (13773–1-AP) were from Proteintech (IL, USA); phospho-PKCδ (Y311) (ab76181), PKCdelta (ab182126), SHP-1 (ab227503), HA (ab9110) were from Abcam (MA, USA); BIP (M010300), MyD88 (P012343) were from Epizyme, Shanghai, China); and ATF4 (11815), sXBP-1 (40435) were from Cell Signaling Technology (MA, USA).

Techniques: Migration, Transwell Assay, Cell Culture, Transfection, Staining, Immunohistochemistry, Co-Culture Assay

Journal: eBioMedicine

Article Title: USP25 regulates atherosclerosis by restricting RIPK1-mediated inflammatory responses

doi: 10.1016/j.ebiom.2026.106213

Figure Lengend Snippet: USP25 is downregulated in human and mouse atherosclerotic lesions. (A) Screening of USP family genes in human carotid artery plaques based on GSE100927 and GSE41571 datasets. In volcano plots, red and blue dots represent upregulated and downregulated USPs, respectively. (B) Transcriptional levels of selected USPs in aortas of ApoE −/− mice fed with ND and HFD were determined by qRT-PCR. ∗P < 0.05, ∗∗P < 0.01, ns, not significant, unpaired two-tailed Student's t- test (n = 5). (C) Representative immunoblots (upper panel) and densitometric quantification (lower panel) of USP25 in aortas of ND and HFD-fed ApoE −/− mice. ∗∗P < 0.01, unpaired two-tailed Student's t- test (n = 3). (D) Representative immunoblots (upper panel) and densitometric quantification (lower panel) of USP25 in mild and severe atherosclerotic lesions of human carotid arteries. ∗P < 0.05, unpaired two-tailed Student's t- test (n = 3). (E–G) Representative immunofluorescence staining of USP25 and F4/80 (E), α-SMA (F), as well as CD31 (G) in aortic roots. Scale bar: 15 μm. (H) Representative immunofluorescence staining of USP25 and F4/80 in aortas of ApoE −/− mice fed with HFD for indicated time. Scale bar: 100 μm ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, two-way ANOVA with Bonferroni post hoc test (n = 5). (I) Representative immunofluorescence staining of USP25 and CD68 in carotid arteries from patients with mild and severe atherosclerosis. Scale bar: 100 μm.

Article Snippet: Antibodies against F4/80 (Cat#: sc-377009, 1: 50 for immunofluorescence, RRID: AB_2927461 ) and USP47 (Cat#: sc-100633, 1:500, RRID: AB_2241454 ) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Quantitative RT-PCR, Two Tailed Test, Western Blot, Immunofluorescence, Staining

Journal: eBioMedicine

Article Title: USP25 regulates atherosclerosis by restricting RIPK1-mediated inflammatory responses

doi: 10.1016/j.ebiom.2026.106213

Figure Lengend Snippet: USP25 deficiency exacerbates atherosclerosis in HFD-fed ApoE −/− mice. (A) Representative Oil Red O staining of aortas from HFD-fed Usp25 +/+ ApoE −/− and Usp25 −/− ApoE −/− mice. Scale bar: 5 mm. (B) Data show the percentage of plaque area/total vessel area. AA, aortic arch; TA, thoracic aorta. ∗∗∗P < 0.001, unpaired 2-tailed Student's t test (n = 6). (C) Representative H&E staining of aortic root sections. Scale bar: 200 μm. (D) Quantification of lesion area (left) and percentages of necrotic core (right). ∗∗P < 0.01, unpaired 2-tailed Student's t test (n = 10). (E) Representative Oil Red O (top), anti-F4/80 immunofluorescence (middle), and anti-α-SMA immunofluorescence (bottom) staining of aortic root sections. Scale bar: 100 μm. (F) Percentages of Oil Red O (top), F4/80 + (middle), and α-SMA + (bottom) area in aortic root sections. ∗∗P < 0.01 , ∗∗∗P < 0.001, unpaired 2-tailed Student's t test (n = 5). (G) Protein levels of IL-6 and TNF-α in atherosclerotic lesions were analysed using ELISA. ∗∗P < 0.01 , ∗∗∗P < 0.001, unpaired 2-tailed Student's t test (n = 6). (H) Transcriptional levels of indicated genes in aortas were determined by qRT-PCR. Target gene expression was normalized to the level of Actb mRNA. ∗P < 0.05, ∗∗P < 0.01, unpaired 2-tailed Student's t test (n = 5).

Article Snippet: Antibodies against F4/80 (Cat#: sc-377009, 1: 50 for immunofluorescence, RRID: AB_2927461 ) and USP47 (Cat#: sc-100633, 1:500, RRID: AB_2241454 ) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Staining, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Targeted Gene Expression

Journal: eBioMedicine

Article Title: USP25 regulates atherosclerosis by restricting RIPK1-mediated inflammatory responses

doi: 10.1016/j.ebiom.2026.106213

Figure Lengend Snippet: Deficiency of USP25 in hematopoietic cells exacerbates atherosclerosis in HFD-fed ApoE −/− mice. (A) Representative Oil Red O staining of aortas from Usp25 +/+ → ApoE −/− and Usp25 −/− → ApoE −/− mice fed a HFD for 16 weeks. Scale bar: 5 mm. (B) Data show the percentage of plaque area/total vessel area. ∗∗∗P < 0.001, unpaired two-tailed Student's t test (n = 6). (C) H&E staining of aortic root sections. Scale bar: 200 μm. (D) Quantitative analysis of lesion area (left) and percentages of necrotic cores (right). ∗∗ P < 0.01, unpaired two-tailed Student's t test (n = 10). (E) Representative Oil Red O (top), anti-F4/80 immunofluorescence (middle), and anti-α-SMA immunofluorescence (bottom) staining of aortic root sections. Scale bar: 100 μm. (F) Percentages of Oil Red O (top), F4/80 + (middle), and α-SMA + (bottom) area in aortic root sections. ∗∗P < 0.01, unpaired two-tailed Student's t test (n = 5). (G) Protein levels of IL-6 and TNF-α in atherosclerotic lesions were analysed using ELISA. ∗∗P < 0.01 , ∗∗∗P < 0.001, unpaired 2-tailed Student's t test (n = 6). (H) Aortas were isolated from Usp25 +/+ → ApoE −/− and Usp25 −/− → ApoE −/− mice after 16 weeks of HFD feeding. Transcriptional levels of indicated genes in aortas were determined by qRT-PCR. ∗∗P < 0.01, ∗∗∗P < 0.001, unpaired two-tailed Student's t test (n = 6).

Article Snippet: Antibodies against F4/80 (Cat#: sc-377009, 1: 50 for immunofluorescence, RRID: AB_2927461 ) and USP47 (Cat#: sc-100633, 1:500, RRID: AB_2241454 ) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Staining, Two Tailed Test, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Isolation, Quantitative RT-PCR

Journal: eBioMedicine

Article Title: USP25 regulates atherosclerosis by restricting RIPK1-mediated inflammatory responses

doi: 10.1016/j.ebiom.2026.106213

Figure Lengend Snippet: USP25 deficiency increases inflammatory responses in macrophages by enhancing NF-κB activation. (A) BMDMs isolated from Usp25 +/+ and Usp25 −/− mice were stimulated with or without ox-LDL (50 μg/mL) for 6 h. The transcriptional levels of the indicated genes were detected by qRT-PCR. ∗P < 0.05, ∗∗P < 0.01, two-way ANOVA with Bonferroni post hoc test (n = 3). (B) BMDMs isolated from Usp25 +/+ and Usp25 −/− mice were incubated with or without ox-LDL (50 μg/mL) for 24 h, followed by Oil Red O staining. Scale bar: 20 μm. (C) Quantitation of Oil Red O-positive areas in BMDMs. ∗∗∗P < 0.001, two-way ANOVA using Bonferroni's post hoc test (n = 5). (D) KEGG pathway enrichment analysis of signalling pathways increased in macrophages of human atherosclerotic lesions based on GSE159677 . (E) BMDMs from Usp25 +/+ and Usp25 −/− mice were treated with or without ox-LDL (50 μg/mL) for 30 min, followed by Western blot analysis with indicated antibodies. (F–G) Representative immunofluorescence staining (F) and quantitation (G) of p65 nuclear translocation in BMDMs treated with or without ox-LDL (50 μg/mL). Scale bar: 50 μm ∗∗∗P < 0.001, two-way ANOVA with Bonferroni post hoc test (n = 3). (H–I) Representative immunofluorescence staining (H) and quantification (I) of p-p65 and F4/80 in aortic roots of HFD-fed Usp25 +/+ ApoE −/− and Usp25 −/− ApoE −/− mice. Scale bar: 50 μm ∗∗∗P < 0.001, unpaired two-tailed Student's t test (n = 6).

Article Snippet: Antibodies against F4/80 (Cat#: sc-377009, 1: 50 for immunofluorescence, RRID: AB_2927461 ) and USP47 (Cat#: sc-100633, 1:500, RRID: AB_2241454 ) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Activation Assay, Isolation, Quantitative RT-PCR, Incubation, Staining, Quantitation Assay, Western Blot, Immunofluorescence, Translocation Assay, Two Tailed Test

Journal: eBioMedicine

Article Title: USP25 regulates atherosclerosis by restricting RIPK1-mediated inflammatory responses

doi: 10.1016/j.ebiom.2026.106213

Figure Lengend Snippet: USP25 affects atherosclerosis by regulating RIPK1. (A) Experimental flowchart for the establishment of atherosclerosis in AAV-infected bone marrow chimeric mice. (B) Representative Oil Red O staining of aortas from indicated mice fed a HFD for 16 weeks. Scale bar: 5 mm. (C) Data show the percentage of plaque area/total vessel area. ∗∗∗P < 0.001, unpaired two-tailed Student's t test (n = 6). (D) H&E staining of aortic root sections. Scale bar: 200 μm. (E) Quantification of lesion area (left) and percentages of necrotic core (right). ∗∗∗P < 0.001, unpaired 2-tailed Student's t test (n = 10). (F) Representative Oil Red O staining of aortic root sections. Scale bar: 100 μm. (G) Percentages of Oil Red O area in aortic root sections. ∗∗P < 0.01, unpaired two-tailed Student's t test (n = 5). (H–I) Representative immunofluorescence staining (H) and quantification (I) of F4/80 in aortic root sections. Scale bar: 100 μm ∗∗∗P < 0.001, unpaired two-tailed Student's t test (n = 5). (J) Protein levels of TNF-α and IL-6 in atherosclerotic lesions were analysed using ELISA. ∗∗P < 0.01, unpaired two-tailed Student's t test (n = 6). (K) Transcriptional levels of indicated genes in aortas were determined by qRT-PCR. ∗P < 0.05, ∗∗P < 0.01, unpaired two-tailed Student's t test (n = 5).

Article Snippet: Antibodies against F4/80 (Cat#: sc-377009, 1: 50 for immunofluorescence, RRID: AB_2927461 ) and USP47 (Cat#: sc-100633, 1:500, RRID: AB_2241454 ) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Infection, Staining, Two Tailed Test, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: eBioMedicine

Article Title: USP25 regulates atherosclerosis by restricting RIPK1-mediated inflammatory responses

doi: 10.1016/j.ebiom.2026.106213

Figure Lengend Snippet: USP25 overexpression alleviates atherosclerosis in HFD-fed ApoE −/− mice. (A) Representative Oil Red O staining of aortas from Usp25 +/+ → ApoE −/− and Usp25 Tg → ApoE −/− mice fed a HFD for 16 weeks. Scale bar: 5 mm. (B) Data show the percentage of plaque area/total vessel area. ∗∗∗P < 0.001, unpaired two-tailed Student's t test (n = 6). (C) Aortas from HFD-fed Usp25 +/+ → ApoE −/− and Usp25 Tg → ApoE −/− mice were lysed for protein isolation. The lysates were immunoprecipitated with anti-RIPK1 antibody, followed by Western blot analysis with indicated antibodies. (D) H&E staining of aortic root sections. Scale bar: 200 μm. (E) Quantification of lesion area (left) and percentages of necrotic core (right). ∗∗P < 0.01, unpaired 2-tailed Student's t test (n = 10). (F) Representative Oil Red O staining of aortic root sections. Scale bar: 100 μm. (G) Percentages of Oil Red O area in aortic root sections. ∗∗P < 0.01, unpaired two-tailed Student's t test (n = 5). (H–I) Representative immunofluorescence staining (H) and quantification (I) of F4/80 in aortic root sections. Scale bar: 100 μm ∗∗P < 0.01, unpaired two-tailed Student's t test (n = 5). (J) Protein levels of TNF-α and IL-6 in atherosclerotic lesions were analysed using ELISA, ∗∗P < 0.01, unpaired two-tailed Student's t test (n = 6). (K) Transcriptional levels of indicated genes in aortas were determined by qRT-PCR. ∗P < 0.05, ∗∗P < 0.01, unpaired two-tailed Student's t test (n = 5).

Article Snippet: Antibodies against F4/80 (Cat#: sc-377009, 1: 50 for immunofluorescence, RRID: AB_2927461 ) and USP47 (Cat#: sc-100633, 1:500, RRID: AB_2241454 ) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Over Expression, Staining, Two Tailed Test, Isolation, Immunoprecipitation, Western Blot, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR