Journal: Nature Communications

Article Title: Perivascular mesenchymal cells instruct ST2+ reparative macrophages to promote endovascular injury-induced neointimal hyperplasia in mice

doi: 10.1038/s41467-026-68587-x

Figure Lengend Snippet: a The experimental design. BMDMs treated by IL-33 (20 ng/mL) for 24 h before additional 24 h fresh medium incubation to prepare CMs. b Representative images and quantification of EdU+ VSMCs co-cultured with the indicated CMs. Scale bar, 100 μm. c Analysis of the putative ligand-receptor communications between arterial macrophages and VSMCs based on the scRNA-seq data of D0 and D7 using CellChat. d Volcano plot depicting the upregulated and downregulated genes in macrophages at D0 and D7. e Violin plots showing Spp1 gene expression of the 7 immune cell populations based on scRNA-seq data. f (Left) Representative flow cytometry gating for quantifying OPN-expressing macrophages and neutrophils at D7. (Right) OPN+ cell percentages and OPN signal intensities in macrophages and neutrophils at D7. n = 6 mice per group. g Representative images of immunofluorescence staining of MAC2 and OPN using the artery sections from mice at D7. Scale bars, 50 μm. h mRNA levels of Spp1 in the artery from mice at D0 ( n = 4), D2 ( n = 8), D4 ( n = 8), D7 ( n = 8) and D14 (n = 4). i Plasma OPN protein levels in patients with or without in-stent restenosis (ISR, n = 25; Non-ISR, n = 25). j mRNA levels of Spp1 of the artery from Control ( n = 7) and Pb- Il33 KO ( n = 8) at D7. k mRNA levels of Spp1 in the artery from Control ( n = 10) and Mac- Il1rl1 KO ( n = 7) at D7. l mRNA levels of Spp1 in IL-33-treated (20 ng/mL) BMDMs for 24 h. m BMDMs electro-transfected with si Spp1 followed by IL-33 (20 ng/mL) treatment to prepare CMs. n mRNA levels of the cell proliferation-related genes in indicated CM-exposed VSMCs. o Representative H&E staining images and Intima/Media ratios and lumen diameter/aspect ratios in the artery sections at D28. Mice post-FAI were administrated with the hydrogel containing si Spp1 or si NC locally at the injured sites. n = 6 mice for each group. Scale bars, 50 μm. Data are all shown as the mean ± s.e.m. ** p < 0.01 or *** p < 0.001 by unpaired two-tailed Student’s t -test ( f , i , j , k , n , o ), one-way ( b , h ) or two-way ANOVA ( l ). Source data and statistic information are provided as Source data files. Figure 7a,c, m were created in BioRender. Shan, B. (2025) https://BioRender.com/x16x8wd .

Article Snippet: Plasma IL-33 and OPN protein levels were determined by a commercially available human IL33 ELISA Kit (#EK133, Multi sciences) and human OPN ELISA Kit (#EK1135, Multi Sciences) according to the manufacturer’s instructions.

Techniques: Incubation, Cell Culture, Gene Expression, Flow Cytometry, Expressing, Immunofluorescence, Staining, Clinical Proteomics, Control, Transfection, Two Tailed Test

Journal: Nature Communications

Article Title: Single-cell multiomics uncovers an endothelial mechanosensitive PIEZO1-IL-33 axis driving pulmonary fibrosis

doi: 10.1038/s41467-026-70193-w

Figure Lengend Snippet: A CellChat analysis of human lung snRNA-seq data. B Venn of 47 shared DEGs from four datasets ( GSE159354 , GSE132771 , GSE128033 , GSE122960 ). C Heatmap of 47-gene expression across cell types. D Correlation analysis (Pearson Correlation) of expression of IGFBP5, IL33, MGP, SPRY1, ACKR1 and mechanical stress scores in ECs. E Correlation analysis (Pearson Correlation) of expression of PIEZO1 and IGFBP5, IL33, MGP, SPRY1, ACKR1 in ECs. F Human snRNA-seq UMAP and violin of IL33 distribution and subpopulations. G Bar chart showed expression of IL33 (mean ± SEM) in IPF (n = 7966 cells) vs control (n = 4174 cells). P value was calculated using the two-tailed unpaired t-test. H Violin of IL33 in PIEZO1 - vs PIEZO1 + ECs. I Bulk-RNA-seq of saline and BLM (7 days and 14 days) induced mice (n = 3 per group). Gene expression levels were Z-score normalized. P value was calculated using the two-tailed unpaired t-test. J qPCR of Il33 in BLM mouse lung ECs. K IL-33 (green) and CD31 (red) co-staining in human IPF vs normal, quantification is shown at right (n = 10). L IL-33 (green) and CD31 (red) co-staining in Yoda1- or GsMTx4-treated fibrotic mouse lungs, quantification is shown at right (n = 3). M IL-33 (green) and CD31 (red)co-staining in Piezo1 WT and Piezo1 ΔEC BLM lungs, quantification is shown at right (n = 3). Error bars represent mean ± SEM, H, K, J, M: two-tailed unpaired t-test; L: one-way ANOVA followed Tukey’s multiple comparwasons test, All n values indicate biologically independent samples. Source data are provided as a Source Data file.

Article Snippet: The amounts of IL-33 in HUVECs supernatant were quantified using the Human IL-33 DuoSet ELISA kit (R&D Systems, #D3300B) according to the manufacturer’s instructions.

Techniques: Gene Expression, Expressing, Control, Two Tailed Test, RNA Sequencing, Saline, Staining

Journal: Nature Communications

Article Title: Single-cell multiomics uncovers an endothelial mechanosensitive PIEZO1-IL-33 axis driving pulmonary fibrosis

doi: 10.1038/s41467-026-70193-w

Figure Lengend Snippet: A Schematic of BLM-induced fibrosis in Il33 WT vs Il33 ΔEC mice. B Il33 mRNA (n = 4) and IL-33 protein in lung ECs (n = 3). C Representative Masson/PSR images and quantification (n = 5). D αSMA immunofluorescence and quantification (n = 4). E Lung hydroxyproline content (n = 5). F Schematic: EC-specific Il33 overexpression via AAV-Tie1 in Piezo1 WT and Piezo1 ΔEC mice; 4-week pretreatment, BLM at week 4, sacrifice at week 7. G IL-33 expression in ECs, immune and epithelial cells after AAV-OE- Il33 (n = 3). H Masson/PSR images and quantification (n = 5). I αSMA immunofluorescence and quantification (n = 4). J Hydroxyproline content (n = 5). Error bars: mean ± SEM; B,G: two-tailed unpaired t-test; C,D-E,I: two-way ANOVA followed Tukey’s multiple comparisons test; H,J: three-way ANOVA followed Tukey’s multiple comparisons test. All n values indicate biologically independent samples. Source data are provided as a Source Data file.

Article Snippet: The amounts of IL-33 in HUVECs supernatant were quantified using the Human IL-33 DuoSet ELISA kit (R&D Systems, #D3300B) according to the manufacturer’s instructions.

Techniques: Immunofluorescence, Over Expression, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: Single-cell multiomics uncovers an endothelial mechanosensitive PIEZO1-IL-33 axis driving pulmonary fibrosis

doi: 10.1038/s41467-026-70193-w

Figure Lengend Snippet: HUVECs under 20% stretch for 0,6,24 h or 2,12,25 kPa substrates; IL-33 secretion ( n = 3) A and mRNA (n = 3) B . C PIEZO1 knock-down with sh PIEZO1 ( n = 3). HUVECs were transduced with sh PIEZO1 for 48 h, cultured on 25 kPa substrates for 24 h or subjected to 6 h mechanical stretch before measuring IL-33 secretion ( n = 3) D and mRNA ( n = 3) E . HUVECs were cultured on 2, 12, 25 kPa substrates for 24 h or stretched (20%) for 0, 6, 24 h, followed by measurement of calpain activity F and CAPN2 protein G ( n = 3). HUVECs (sh PIEZO1 ) were cultured on 25 kPa substrates for 24 h or subjected to 6 h mechanical stretch, followed by measurement of calpain activity ( n = 3) H and CAPN2 protein levels ( n = 3) I . J CAPN2 knock-down with shCAPN2 ( n = 3). K HUVECs (sh CAPN2 ) were cultured on 25 kPa substrates for 24 h or subjected to 6 h mechanical stretch, IL-33 secretion(left) and mRNA(right) ( n = 3). L Model for paracrine IL-33 regulation. M Cistrome: STAT3 predicted to drive IL-33 regulation. N intersection of top 100 motifs in BLM-treated mouse ECs with top 10 TFs. O The protein levels of P-STAT3 and STAT3 in HUVECs treated with 20% stretch for 0 and 6 hours ( n = 3). P The protein levels of P-STAT3 and STAT3 in HUVECs (sh PIEZO1 or sh CAPN2 ) treated with 20% stretch for 6 h ( n = 3). Q STAT3 knock-down with sh STAT3 ( n = 3). R HUVECs (sh STAT3 ) were cultured on 25 kPa substrates for 24 h or subjected to 6 h mechanical stretch; IL-33 secretion(left) and mRNA(right) ( n = 3). Error bars: mean ± SEM; C , G , J , O – Q : two-tailed unpaired t-test; A , B , F : one-way ANOVA followed Šídák’s multiple comparisons test, D – E , H – I , K , R : two-way ANOVA Tukey’s multiple comparisons test. All n denote biologically independent samples. Source data are provided as a Source Data file.

Article Snippet: The amounts of IL-33 in HUVECs supernatant were quantified using the Human IL-33 DuoSet ELISA kit (R&D Systems, #D3300B) according to the manufacturer’s instructions.

Techniques: Knockdown, Transduction, Cell Culture, Activity Assay, Two Tailed Test

Journal: Molecular Medicine

Article Title: Dual and context-dependent role of the interleukin-33/soluble suppression of tumorigenicity 2 axis in obesity and adipose tissue inflammation

doi: 10.1186/s10020-026-01454-z

Figure Lengend Snippet: Effect of obesity and sex on the circulating levels of interleukin (IL)-33 and soluble suppression of tumorigenicity 2 (sST2). Fasting serum concentrations of ( A ) IL-33, ( C ) sST2 and ( E ) IL-33/sST2 in normal-weight (NW-NG) volunteers ( n = 22), patients with obesity with normoglycemia (OB-NG) ( n = 24) and with obesity-associated type 2 diabetes (OB-T2D) ( n = 27). Circulating concentrations of ( B ) IL-33, ( D ) sST2 and ( F ) IL-33/sST2 in males ( n = 26) and females ( n = 47) volunteers. Data are mean ± SEM. Differences between groups were analyzed by one-way ANOVA followed by Tukey’s tests or by unpaired two-tailed Student’s t test as appropriate. * P < 0.05, ** P < 0.01 and *** P < 0.001

Article Snippet: Circulating concentrations of IL-33 (D3300B) and sST2 (DST200) were quantified using commercial ELISA kits (R&D Systems, Minneapolis, MN) according to manufacturer’s instructions.

Techniques: Two Tailed Test

Journal: Molecular Medicine

Article Title: Dual and context-dependent role of the interleukin-33/soluble suppression of tumorigenicity 2 axis in obesity and adipose tissue inflammation

doi: 10.1186/s10020-026-01454-z

Figure Lengend Snippet: Gene and protein expression levels of IL-33 in visceral adipose tissue (VAT). A Bar graphs show the gene expression levels of IL33 in VAT samples from volunteers with normal weight (NW-NG, n = 8) and people with obesity and normoglycemia (OB-NG, n = 13) and with obesity-associated type 2 diabetes (OB-T2D, n = 16). B Protein expression levels of IL-33 in VAT from volunteers with normal weight (NW-NG, n = 4) and people with obesity and normoglycemia (OB-NG, n = 13) and with obesity-associated type 2 diabetes (OB-T2D, n = 10). C Scatter plots show the correlations between mRNA expression and protein levels of IL-33 as well as their associations with body fat percentage (BF). Correlation coefficients (r) and corresponding P values were calculated using Pearson’s correlation test. D Representative immunostaining for IL-33 in VAT samples from people with OB-NG and with OB-T2D (magnification 200x). E Comparison of gene expression levels of IL33 with IL1A , IL1B , IL4 , IL6 , IL8 , IL10 , IL13 , IL18 and IL32 in VAT obtained from people with obesity. Values are the mean ± SEM. Differences between groups were analyzed by one-way ANOVA followed by Tukey’s tests or by unpaired two-tailed Student’s t test as appropriate. * P < 0.05, and ** P < 0.01

Article Snippet: Circulating concentrations of IL-33 (D3300B) and sST2 (DST200) were quantified using commercial ELISA kits (R&D Systems, Minneapolis, MN) according to manufacturer’s instructions.

Techniques: Expressing, Gene Expression, Immunostaining, Comparison, Two Tailed Test

Journal: Molecular Medicine

Article Title: Dual and context-dependent role of the interleukin-33/soluble suppression of tumorigenicity 2 axis in obesity and adipose tissue inflammation

doi: 10.1186/s10020-026-01454-z

Figure Lengend Snippet: Gene expression levels of inflammation-related factors in human visceral adipocytes treated with different concentrations of ( A ) IL-33 and ( B ) LPS for 3 h followed by IL-33 for another 24 h. Values are the mean ± SEM ( n = 6 per group). Differences between groups were analyzed by one-way ANOVA followed by Tukey’s tests. ** P < 0.01. ADIPOQ, adiponectin; IL , interleukin; ITLN1 , omentin; LPS , lipopolysaccharide; TNF , tumor necrosis factor α

Article Snippet: Circulating concentrations of IL-33 (D3300B) and sST2 (DST200) were quantified using commercial ELISA kits (R&D Systems, Minneapolis, MN) according to manufacturer’s instructions.

Techniques: Gene Expression

Journal: Molecular Medicine

Article Title: Dual and context-dependent role of the interleukin-33/soluble suppression of tumorigenicity 2 axis in obesity and adipose tissue inflammation

doi: 10.1186/s10020-026-01454-z

Figure Lengend Snippet: A Effects of adipocyte-conditioned media (ACM) in monocytes-derived macrophages. Bar graphs show the effect of ACM (20 and 40%) from subjects with obesity incubated for 24 h on the transcript levels of IL33 . Impact of IL-33 on the expression levels of inflammation-related factors in monocytes-derived macrophages primed with ( B ) ACM and ( C ) LPS. Values are the mean ± SEM ( n = 6 per group). Differences between groups were analyzed by one-way ANOVA followed by Tukey’s tests. * P < 0.05 and ** P < 0.01. CCL2 , monocyte chemoattractant protein-1; IL , interleukin; LPS , lipopolysaccharide; TNF , tumor necrosis factor-α

Article Snippet: Circulating concentrations of IL-33 (D3300B) and sST2 (DST200) were quantified using commercial ELISA kits (R&D Systems, Minneapolis, MN) according to manufacturer’s instructions.

Techniques: Derivative Assay, Incubation, Expressing

Journal: Molecular Medicine

Article Title: Dual and context-dependent role of the interleukin-33/soluble suppression of tumorigenicity 2 axis in obesity and adipose tissue inflammation

doi: 10.1186/s10020-026-01454-z

Figure Lengend Snippet: The increased circulating levels of IL-33 in obesity together the decrease in its soluble decoy receptor sST2 may be potentially enhancing IL-33 bioactivity. In addition, gene and protein expression levels of IL-33 in visceral adipose tissue (VAT) are upregulated in obesity in both adipocytes and stromal vascular fraction cells. IL-33 modulates adipocyte function by upregulating both anti-inflammatory mediators ( ADIPOQ , ITLN1 and IL13 ) and pro-inflammatory cytokines ( TNF , IL8 ). Under inflammatory conditions, such as LPS stimulation, IL-33 attenuates excessive pro-inflammatory responses and promotes protective adipokine expression in visceral adipocytes. In THP-1-derived macrophages, adipocyte-derived factors suppress IL33 expression, whereas IL-33 attenuates macrophage pro-inflammatory activation and enhances IL4 expression only in the context of adipocyte-conditioned medium. Together, these findings suggest the dual and context-dependent nature of IL-33 in obesity, acting as a compensatory mediator to maintain tissue homeostasis while potentially contributing to chronic low-grade inflammation when elevated chronically. ADIPOQ, adiponectin; IL , interleukin; ITLN1 , omentin; TNF , tumor necrosis factor α

Article Snippet: Circulating concentrations of IL-33 (D3300B) and sST2 (DST200) were quantified using commercial ELISA kits (R&D Systems, Minneapolis, MN) according to manufacturer’s instructions.

Techniques: Expressing, Derivative Assay, Activation Assay