Journal: Nature Immunology

Article Title: The IL-22–oncostatin M axis promotes intestinal inflammation and tumorigenesis

doi: 10.1038/s41590-025-02149-z

Figure Lengend Snippet: a , H. hepaticus + anti-IL-10R colitis was induced in wild-type mice for 14 days. Protein analysis of IECs from steady-state ( n = 4) and H. hepaticus + anti-IL-10R ( n = 5) colitic mice was conducted. The western blot depicts phosphorylated STAT1 (pSTAT1), STAT1, pSTAT3, STAT3 and β-actin for representative samples from two experiments. b , c , Immunofluorescence staining for pSTAT1 and pSTAT3 in steady-state ( n = 7) and inflamed ( n = 10) mouse samples ( b ) with subsequent quantification ( c ) in the epithelium; scale bar, 20 μm. Hpf, high-power field. Data were pooled from two experiments. d , e , H. hepaticus + anti-IL-10R colitis was induced in wild-type mice for 7 days, and mice were treated with anti-IL-22 (clone 8e11, n = 4) or isotype control (mouse IgG1, GP120:9709, n = 5) on days 0 and 3 of colitis induction. Protein analysis of IECs from steady-state and H. hepaticus + anti-IL-10R colitic mice and anti-IL-22 or isotype-treated inflamed mice was performed. d , Western blot depicting STAT3 phosphorylation in epithelial cells from anti-IL-22- or isotype-treated colitic mice. Data are representative of two independent experiments. e , Quantification of STAT3 phosphorylation using ImageJ. The relative band intensity of blots from inflamed mice was normalized to that of steady-state untreated mice. f , g , Colitis was induced in Vil creERT2 Stat3 fl/fl (IEC Δ Stat3 ) or Vil creERT2 Stat3 fl/wt (IEC WT ; f ) and Stat1 –/– or Stat1 +/+ ( g ) mice using the H. hepaticus + anti-IL-10R model for 7 days. qPCR analysis was performed to assess the expression of Osmr and Reg3g in epithelial cells from inflamed mice normalized to epithelial cells isolated from untreated respective control mice. Data are representative of one experiment; n = 5–7 mice per genotype. h , Mouse colon organoids generated from Vil cre+ Stat3 fl/fl (IEC Δ Stat3 ) or Vil cre- Stat3 fl/fl (IEC WT ) mice and stimulated with 10 ng ml –1 IL-22. Relative expression of Osmr was analyzed by qPCR. Data are representative of three independent experiments from three independent biological replicates. P values (two-tailed) were calculated using a Mann–Whitney test for c and e – h .

Article Snippet: HCA-7 cells were also stimulated with 100 ng ml –1 recombinant human IL-22 or recombinant human OSM for 30 min, 24 h or 48 h, followed by analysis via MSD pSTAT3 assay (Meso Scale Diagnostics).

Techniques: Western Blot, Immunofluorescence, Staining, Control, Phospho-proteomics, Expressing, Isolation, Generated, Two Tailed Test, MANN-WHITNEY

Journal: Nature Immunology

Article Title: The IL-22–oncostatin M axis promotes intestinal inflammation and tumorigenesis

doi: 10.1038/s41590-025-02149-z

Figure Lengend Snippet: ( a ) Immunofluorescence staining for pSTAT1 and pSTAT3 in steady-state (n = 7) and inflamed (n = 10) mouse samples with subsequent quantification in colon lamina propria. Data pooled from two experiments. P -values are from Mann-Whitney U tests. ( b ) Western blot analysis of pSTAT3, total STAT3, and β-actin in mouse colon organoids treated with the indicated cytokines. Data are representative of two independent experiments. ( c ) Western blot analysis of pSTAT1, total STAT1, and β-actin in mouse colon organoids treated with indicated cytokines. Data are representative of two independent experiments. ( d ) Western blot analysis of pSTAT1, total STAT1, and β-actin in the human Caco2 cell line (left panel) was performed to confirm functionality of pSTAT1 antibody. Epithelial cells from C57BL/6 J mice with induced inflammation at days 7 and 14 (n = 5) were also analyzed (right panel). pSTAT1, total STAT1, and β-actin in steady state epithelial cells from C57BL/6 J mice are shown in Fig. . ( e ) Colon epithelial organoids isolated from C57BL/6 mice were treated with JAK/STAT pathway inhibitors (50 μM fludarabine, 25 μM STA-21, 2.5 μM ruxolitinib, or 2.5 μM tofacitinib) for 6 h, and 10 ng/mL of IL-22 was added for another 6 h. 0.1% DMSO was used as inhibitor control. Relative expression of Osmr was analyzed by qPCR. Data are shown for n = 2 biological replicates with n = 3 technical replicates. Graph displays mean ± SEM. P -values are derived from Kruskal-Wallis test.

Article Snippet: HCA-7 cells were also stimulated with 100 ng ml –1 recombinant human IL-22 or recombinant human OSM for 30 min, 24 h or 48 h, followed by analysis via MSD pSTAT3 assay (Meso Scale Diagnostics).

Techniques: Immunofluorescence, Staining, MANN-WHITNEY, Western Blot, Isolation, Control, Expressing, Derivative Assay

Journal: Nature Immunology

Article Title: The IL-22–oncostatin M axis promotes intestinal inflammation and tumorigenesis

doi: 10.1038/s41590-025-02149-z

Figure Lengend Snippet: ( a ) Human HCA-7 colon epithelial cells were stimulated with 100 ng/ml of rhIL-22 or rhOSM for 30 min, 24 or 48 h. Subsequently, pSTAT3 was quantified in cell lysates using an electrochemiluminescence assay (MSD). Data are representative of two independent experiments, each with three technical replicates. P -values derived from a 2-way comparison using Tukey’s multiple comparison test. ( b ) Analysis of STAT3 phosphorylation after colitis induction in IECs from IEC Δ Osmr and control mice (values normalized to those from steady-state mice). P -values are derived from Mann–Whitney U test. Data are representative of two independent experiments; n = 6 mice per group. ( c ) Volcano plot depicting differentially expressed genes in the epithelial cell line, HCA-7, after treatment with rhOSM for 24 h as described in ( a ). ( d ) GSEA plot depicting enrichment of OSM-activated genes in epithelial cells (defined in analysis shown in panel c ) comparing inflamed IEC Δ Osmr and control mice, as in Fig. .

Article Snippet: HCA-7 cells were also stimulated with 100 ng ml –1 recombinant human IL-22 or recombinant human OSM for 30 min, 24 h or 48 h, followed by analysis via MSD pSTAT3 assay (Meso Scale Diagnostics).

Techniques: Electrochemiluminescence, Derivative Assay, Comparison, Phospho-proteomics, Control, MANN-WHITNEY

Journal: Nature Immunology

Article Title: The IL-22–oncostatin M axis promotes intestinal inflammation and tumorigenesis

doi: 10.1038/s41590-025-02149-z

Figure Lengend Snippet: a , Wild-type mice underwent AOM/DSS-induced CAC. Colon tissues from tumor-bearing mice were analyzed by ISH for Osmr (blue) and Pdgfra (red; stromal marker); scale bars, 1,000 μm (left) and 200 μm (two insets on the right). b , Spatial transcriptomics analysis of a biopsy from an individual with CAC, including tumor tissue (red box) and adjacent normal mucosa (green box). OSM and OSMR expression was assessed in epithelial and nonepithelial cells using epithelial ( EPCAM , CDH1 , KRT20 , MUC5B and MKI67 ) and nonepithelial markers ( CD3E , CD4 , CD14 , CD68 , C1QC , CD19 , CD79A , PDGFRA , VIM , PECAM1 , CD34 and KIT ); scale bar, 1,000 μm (left) and 100 μm (insets on the right). c , Quantification of OSMR expression in epithelial and nonepithelial cells in CAC ( n = 10, red bars) and healthy control ( n = 3, black bars) tissue, along with nonepithelial OSM expression. d , Experimental schematic comparing CAC induction in wild-type versus Osm –/– mice. Representative colonoscopy images and tumor burden on day 80 are shown ( n = 25 per genotype). e , Wild-type mice received AOM and DSS, followed by treatment with anti-OSM ( n = 28) or isotype antibody (rIgG2a; n = 26). Tumor burden, multiplicity and volume were assessed on day 80. Data are shown normalized to the isotype-treated group. f , Immunostaining for pSTAT3 in colon tumors and adjacent normal tissues from mice treated as in e ; scale bar, 200 μm. g , Quantification of pSTAT3 + epithelial cells ( n = 10 per group). h , Relative pSTAT3 expression intensity normalized to that observed in control tissue ( n = 10 per group). Data are derived from two experiments. P values (two-tailed) were calculated from one-sample t -tests and Wilcoxon rank tests. i , j , Vil creERT2+ Osmr fl/fl (IEC Δ Osmr ; n = 15) and Vil creERT2+ Osmr wt/wt (IEC WT ; n = 18) mice were treated with AOM/DSS to induce CAC. Tamoxifen administration induced epithelial-specific Osmr deletion. i , Experimental design and representative colon images. j , Tumor burden, multiplicity and average volumes from two independent experiments. Data are shown normalized to the Vil creERT2+ Osmr wt/wt group. P values (two-tailed) were calculated using a Mann–Whitney test in c – e , g and j .

Article Snippet: HCA-7 cells were also stimulated with 100 ng ml –1 recombinant human IL-22 or recombinant human OSM for 30 min, 24 h or 48 h, followed by analysis via MSD pSTAT3 assay (Meso Scale Diagnostics).

Techniques: Marker, Expressing, Control, Immunostaining, Derivative Assay, Two Tailed Test, MANN-WHITNEY

Journal: PLoS ONE

Article Title: Gender difference in NASH susceptibility: Roles of hepatocyte Ikkβ and Sult1e1

doi: 10.1371/journal.pone.0181052

Figure Lengend Snippet: Wild type (Wt), myeloid IKKβ deficient ( IkbkbΔmye ), or hepatocyte IKKβ deficient ( IkbkbΔhep ) mice in both genders, were fed regular chow (Control) or high cholesterol and saturated fat diet (HCFD) from 2.5 months of age for 20 weeks. (A) Immunoblotting of whole liver lysate and (B) densitometric analysis of pAMPK/AMPK, nSREBP-1c, PPARδ, pJNK-1/JNK-1, and pJNK-2/JNK-2. (C) Plasma adiponectin levels measured by ELISA. (D) Real-time PCR results of PPAR α, δ, and γ genes. *p<0.05, **p<0.01, ***p<0.001 compared to Control diet within gender and within genotype. †p<0.05, ††p<0.01 compared to other genotype within gender and within diet.

Article Snippet: Plasma ALT, insulin and adiponectin concentrations were measured by ELISA kits (ALT: Sigma Diagnostic, St. Louis, MO, insulin: Shibayagi Co., Gunma, Japan, adiponectin: ALPCO Diagnostics, Salem, NH, respectively).

Techniques: Control, Western Blot, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction