Journal: Frontiers in Immunology

Article Title: Sustained STING-IRF7 signaling aggravates LPS-induced endometrial inflammation via excessive neutrophil extracellular traps generation

doi: 10.3389/fimmu.2025.1671848

Figure Lengend Snippet: CD11b was regulated by STING-IRF7 pathway in endometrium. (A) Representative immunohistochemical staining of CD11b in endometrial tissues of WT or STING-deficient mice ( Tmem173 gt ) infected by LPS for 24 h at 400 × magnification (scale bar = 25 μm). (B) The area of CD11b were quantified (n=6). One-way ANOVA test was applied with ** P <0.01 (WT vs. Tmem173 gt in LPS group). (C) STING inhibitor H-151was used to pretreat Ishikawa cell with or without LPS infected for 24 h, then the protein expression of CD11b was detected by Western blotting. (D) Quantification of the amount of CD11b in the four groups (n=5). One-way ANOVA test was applied with * P <0.05 (LPS vs. H-151+LPS group). (E) Quantitative mRNA expression of irf7 in endometrium of mice (n=6). One-way ANOVA test was applied with * P <0.05 (WT vs. Tmem173 gt in LPS group) (F) Representative immunoblots of IRF7 in LPS-stimulated STING-deficient mice ( Tmem173 gt ) endometrium tissues, compared with WT-LPS stimulated mice. (G) Quantification of the amount of IRF7 in the four groups (n=4). One-way ANOVA test was applied with **** P <0.0001(WT vs. Tmem173 gt in LPS group). (H) Representative immunoblots of CEBPB in LPS-stimulated STING-deficient mice ( Tmem173 gt ) endometrium tissues, compared with WT-LPS stimulated mice. (I) Representative immunoblots of IRF7 in STING inhibitor H-151 pretreat Ishikawa cell. (J) Quantification of the amount of IRF7 in the two groups, and t test was applied with * P <0.05 (LPS vs. H-151+LPS group). (K) Quantitative mRNA expression of itgam with IRF7 overexpression in Ishikawa cells, and t test was applied with *** P <0.001 (LPS vs. H-151+LPS group). (L) Representative immunoblots of CD11b with the transfection of IRF7 in Ishikawa cells. (M) Quantification of the amount of CD11b in the two groups, and t test was applied with * P <0.05 (LPS vs. IRF7+LPS group). (N) Detection of CD11b expression by flow cytometry in mice bone marrow neutrophils stimulated with STING inhibitor C-176. (O) Quantification of CD11b expression on Ly6G + CD45 + cells in mice bone marrow neutrophils stimulated with STING inhibitor C-176., and t test was applied with **** P <0.0001. Independent experiments are repeated at least three times.

Article Snippet: Sections were incubated overnight at 4°C with the following primary antibodies diluted in blocking buffer: MPO (1:200, 22225-1-AP, Proteintech, China), CD11b (MAC-1) (1:100, AB133357, Abcam, USA), ELA2 (1:1000, 27642-1-AP, Proteintech, China), Ly-6G (1:100, 87048T; Cell Signaling Technology, USA).

Techniques: Immunohistochemical staining, Staining, Infection, Expressing, Western Blot, Over Expression, Transfection, Flow Cytometry

Journal: bioRxiv

Article Title: TET2 loss promotes premalignant survival and clonal selection in MYC-driven B cell lymphoma

doi: 10.64898/2026.03.20.712678

Figure Lengend Snippet: Kaplan-Meier survival analysis displaying (A) overall survival probability, and (B) median time to death in days for all malignant mice ( EµMyc : n=27, EµMyc Tet2 +/− : n=44, EµMyc Tet2 −/− : n=27). (C) Spleen to body weight ratio in non-malignant (280-320 days) mice ( wildtype : n=7 , Tet2 +/− : n=13 , Tet2 −/− : n=6) and malignant mice, excluding animals without overt disease at 300 days ( EµMyc : n=13, EµMyc Tet2 +/− : n=11, EµMyc Tet2 −/− : n=23). (D) Splenic immune cell composition assessed by flow cytometry: monocytes/macrophages (CD11b + Gr1 − ), granulocytes (CD11b + Gr1 + ), erythroid progenitors (nucleated Ter119 + ), NK cells (TCRβ − NK1.1 + ), CD4 + T cells (TCRβ + CD4 + ), CD8 + T cells (TCRβ + CD8 + ), and malignant B cells (B220 + CD19 + ) ( EµMyc : n=10, EµMyc Tet2 −/− : n=12). (E) Lymphoma immunophenotype in the spleen. Mixed tumors were defined as tumors where neither IgM − nor IgM + cells constituted >80% of the total tumor population. (F) Volcano plots displaying RNA-seq-derived transcriptional profiles of FACS-sorted IgM − tumor (left; B220 + CD19 + IgM − IgD − ) and IgM + tumor (right; B220 + CD19 + IgM + IgD − ) cells. Comparison between EµMyc Tet2 −/− and EµMyc (IgM − tumors: n=4 vs. n=5, IgM + tumors: n=4 vs. n=3) mice was performed separately for each cell type. Significance was defined as adjusted p-value<0.05 and absolute log 2 (fold change)>1. Downregulated genes in EµMyc Tet2 −/− tumors are shown in blue; upregulated genes are shown in red. (G) DNA content and (H) polyploid cell fraction of splenic lymphoma cells, assessed by TO-PRO-3 staining via flow cytometry ( EµMyc : n=7, EµMyc Tet2 −/− : n=8). Flow cytometry assessment of (I) DNA double strand breaks by γH2AX staining ( EµMyc : n=6, EµMyc Tet2 −/− : n=7) and (J) apoptosis by cleaved Caspase-3 staining ( EµMyc : n=5, EµMyc Tet2 −/− : n=5) within splenic lymphoma cells. Bar graphs show median with interquartile range. Statistical significance was determined using (A) Mantel-Cox test, (B, C) one-way ANOVA, (D, G) Mann-Whitney test, or (H, I, J) unpaired t-test, depending on normality (Shapiro-Wilk test) with Holm-Šidák correction for multiple comparisons. ns = not significant, *p<0.05, **p<0.005.

Article Snippet: For depletion of non-B cells, 300 μl of a biotinylated antibody mix (diluted 1:100 in FACS-B) containing αCD4 (Biolegend, 100404), αCD8 (Biolegend, 100704), αNK1.1 (Biolegend, 108704), αCD11b (Milteny Biotec, 130-113-242), αGr1 (Biolegend, 108404) and αTer119 (Biolegend, 116204) was used.

Techniques: Flow Cytometry, RNA Sequencing, Derivative Assay, Comparison, Staining, MANN-WHITNEY

Journal: bioRxiv

Article Title: TET2 loss promotes premalignant survival and clonal selection in MYC-driven B cell lymphoma

doi: 10.64898/2026.03.20.712678

Figure Lengend Snippet: Analysis of splenocytes from premalignant (day 50) EµMyc and EµMyc Tet2 −/− mice. (A) Spleen to body weight ratio for each mouse and (B) absolute splenocyte counts (*10 7 ) ( EµMyc : n=35, EµMyc Tet2 −/− : n=26). (C) Splenic immune cell composition assessed by flow cytometry: monocytes/macrophages (CD11b + Gr1 − ), granulocytes (CD11b + Gr1 + ), erythroid progenitors (nucleated Ter119 + ), NK cells (TCRβ − NK1.1 + ), CD4 + T cells (TCRβ + CD4 + ), CD8 + T cells (TCRβ + CD8 + ), and B cells (B220 + CD19 + ) ( EµMyc : n=18, EµMyc Tet2 −/− : n=13). (D) Splenic B cell subsets were assessed via flow cytometry: IgM − progenitors (B220 + CD19 + IgM − IgD − ), IgM + IgD − immature-(like) (B220 + CD19 + IgM + IgD − ), and IgM + IgD + mature (B220 + CD19 + IgM + IgD + ) B cells. The upper panel summarizes all data ( wildtype : n=10, Tet2 −/− : n=11, EµMyc : n=18, EµMyc Tet2 −/− : n=13), the lower panel shows representative dot blots for the IgM/IgD gate. (E) Volcano plots display RNA-seq-derived transcriptional profiles of premalignant FACS-sorted splenic IgM − progenitors (left; B220 + CD19 + IgM − IgD − ) and IgM + immature(-like) (right; B220 + CD19 + IgM + IgD − ) B cells. Comparisons between EµMyc Tet2 −/− (n=5) and EµMyc (n=6) mice were performed separately for each cell type. Axis ranges were kept identical across volcano plots to allow direct comparison. Significance was defined as adjusted p-value<0.05 and absolute log 2 (fold change)>1. Downregulated genes in EµMyc Tet2 −/− subsets are shown in blue; upregulated genes are shown in red. Bar plots show median with interquartile range. Statistical significance was determined using (A) unpaired t-test or (B, C) Mann-Whitney test and (D) two-way ANOVA, with Holm-Šidák correction for multiple comparisons. Normality was assessed using the Shapiro-Wilk test. n.d. = not detected, ns = not significant, *p<0.05, **p<0.005, ***p<0.0005.

Article Snippet: For depletion of non-B cells, 300 μl of a biotinylated antibody mix (diluted 1:100 in FACS-B) containing αCD4 (Biolegend, 100404), αCD8 (Biolegend, 100704), αNK1.1 (Biolegend, 108704), αCD11b (Milteny Biotec, 130-113-242), αGr1 (Biolegend, 108404) and αTer119 (Biolegend, 116204) was used.

Techniques: Flow Cytometry, RNA Sequencing, Derivative Assay, Comparison, MANN-WHITNEY