Journal: Materials Today Bio

Article Title: Bioinspired lipid droplets nanoplatform for periodontitis therapy: Integrated antibacterial, mitochondrial repair, and immunomodulatory functions

doi: 10.1016/j.mtbio.2026.102808

Figure Lengend Snippet: Verification of the anti-inflammatory activity and polarization-regulating ability of GA@LDs-CRAMP in BMDMs. A) qRT-PCR quantification of inflammatory cytokines ( Il6 , Tnf ), macrophage polarization markers ( Nos2, Arg1 ) and anti-inflammatory cytokine ( Il10 ) (n = 3). B) Quantitative detection of inflammatory cytokine IL-6 (a) and anti-inflammatory cytokine IL-10 (b) in BMDMs culture supernatants by ELISA (n = 5). C) Flow cytometric analysis of the polarization of F4/80-labeled BMDMs macrophages, where CD86 is a marker for M1-type macrophages and CD206 is a marker for M2-type macrophages. D) Quantitative statistical analysis of ( a ) CD86 and (b() b) CD206 fluorescence intensities in cells of each group (n = 3). E) Volcano plot of DEGs between GA@LDs-CRAMP and LPS-treated groups (|log2FC| >1.2, FDR <0.05). F) KEGG pathway enrichment analysis of DEGs (top interested 20 pathways). G) Chord diagram mapping gene-pathway associations for enriched KEGG terms of interest. H) Heatmap of enriched genes of interest. I) GSEA demonstrating enrichment of: (a) cytokine and inflammatory response, (b) oxidative stress and redox regulation, (c) oxidative stress response, (d) TCA cycle, (e) mitochondrial gene expression, (f) oxidative phosphorylation. ns > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: For immunofluorescence staining, sections were incubated with primary antibodies against F4/80 (1:50, Abmart, China, #MU124812) and CD86 (1:100, Santa Cruz, USA, #sc-19617) or CD206 (1:1000, Abcam, USA, #ab64693), FITC-conjugated Goat Anti-Rat IgG (1:100, Proteintech, USA, #SA00003-11), AF647-labeled Goat Anti-Rabbit IgG secondary antibody (1:100), or AF594-labeled Goat Anti-Mouse IgG (1:500, Thermo Fisher, USA, #A-11005) were used respectively.

Techniques: Activity Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Labeling, Marker, Fluorescence, Gene Expression, Phospho-proteomics

Journal: Materials Today Bio

Article Title: Bioinspired lipid droplets nanoplatform for periodontitis therapy: Integrated antibacterial, mitochondrial repair, and immunomodulatory functions

doi: 10.1016/j.mtbio.2026.102808

Figure Lengend Snippet: Immunofluorescence analysis of periodontal tissues. (A) Representative immunofluorescent staining images of F4/80 (green fluorescence) and CD86 (red fluorescence) in periodontal tissues. Scale bar: 100 μm (main), 25 μm (zoom). (B) Quantitative analysis of the percentage of CD86 + F4/80 + double positive cells to F4/80 + single positive cells (n = 5). (C) Representative immunofluorescent staining images of F4/80 (green fluorescence) and CD206 (red fluorescence) in periodontal tissues. Scale bar: 100 μm (main), 25 μm (zoom). (D) Quantitative analysis of the percentage of CD86+F4/80+double positive cells to F4/80+single positive cells (n = 5). ns > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: For immunofluorescence staining, sections were incubated with primary antibodies against F4/80 (1:50, Abmart, China, #MU124812) and CD86 (1:100, Santa Cruz, USA, #sc-19617) or CD206 (1:1000, Abcam, USA, #ab64693), FITC-conjugated Goat Anti-Rat IgG (1:100, Proteintech, USA, #SA00003-11), AF647-labeled Goat Anti-Rabbit IgG secondary antibody (1:100), or AF594-labeled Goat Anti-Mouse IgG (1:500, Thermo Fisher, USA, #A-11005) were used respectively.

Techniques: Immunofluorescence, Staining, Fluorescence

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