Journal: Nature Communications

Article Title: Blocking the m 6 Am methyltransferase PCIF1 releases STAT1-mediated Th1 immunity to potentiate cancer immunotherapy

doi: 10.1038/s41467-026-75269-1

Figure Lengend Snippet: a Schematic of m 6 Am-seq in naïve CD4 + T cells. b Metagene profiles of m 6 Am peak distribution along normalized transcripts. c KEGG pathway enrichment analysis of m 6 Am-modified genes in naïve CD4 + T cells. d Schematic of RiboTag-seq in naïve CD4 + T cells. e Volcano plots of differential translation efficiency (TE) between cKO and WT naïve CD4 + T cells based on RiboTag-seq data. TE was calculated as the ratio of Fragments Per Kilobase of exon model per Million mapped fragments (FPKM) in RiboTag-seq to FPKM in input RNA-seq. f Venn diagram of TE-upregulated genes and m 6 Am-modified genes in naïve CD4 + T cells. g RiboTag-qPCR analysis of Stat1 mRNA association with ribosomes in naïve CD4 + T cells ( n = 4). h–j Representative immunoblot ( h ) and corresponding quantitative data ( i ) ( n = 8), along with representative histograms (left) and quantitative analysis (right) in ( j ) ( n = 6) reveal STAT1 expression in naïve CD4 + T cells from WT or cKO mice. k Schematic (left) and the quantification (right) of the dual-luciferase reporter assay used to characterize the effect of A-to-T substitution at the first nucleotide after the m 7 G cap in the 5’-UTR of Stat1 ( n = 12). l Quantification of IFN-γ expression in Th1-differentiated CD4 + T cells treated with a STAT1 inhibitor (WT or cKO Group n = 6, other Group n = 5). Each n represents biologically independent samples ( k ) or independent samples from individual mouse ( a , e , g – j , l ). Error bars represent means ± SEM; two-tailed, unpaired t -test ( g , i–k ); one-way ANOVA ( l ). p -values are shown on the graphs, and p -values < 0.0001 are reported as such.

Article Snippet: During the first 24 h of Th1 differentiation from naïve CD4 + T cells, the STAT1 inhibitor Fludarabine (2.5 μM; MedChemExpress, 21679-14-1) was added.

Techniques: Modification, RNA Sequencing, Western Blot, Expressing, Luciferase, Reporter Assay, Two Tailed Test

Journal: Journal of Translational Autoimmunity

Article Title: Exploring the immunomodulatory effects of environmental contaminants on autoimmune patients: An in vitro approach

doi: 10.1016/j.jtauto.2025.100341

Figure Lengend Snippet: Modulation of intracellular signaling pathways in HD, RA, and SLE cells exposed to environmental contaminants. Heatmap showing the relative expression and phosphorylation levels of key signaling proteins involved in immune regulation, inflammation, and cell survival (STAT1, STAT3, p38, AKT, and NFκB) and their phosphorylated forms in HD, RA, and SLE after exposure to PM, silica, and TCDD. Data are represented as z-score normalized values. Asterisks indicate statistically significant differences compared with the unstimulated control within each group (p < 0.05). AKT: protein kinase B; HD: healthy donors; NFκB: nuclear factor kappa-light-chain-enhancer of activated B cells; P: phosphorylated form; p38: p38 mitogen-activated protein kinase; PM: particulate matter; RA: rheumatoid arthritis; SLE: systemic lupus erythematosus; STAT: signal transducer and activator of transcription; TCDD: 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Article Snippet: Membranes were stripped and re-probed for total AKT, NFκB p65, p38 MAPK, STAT1, STAT3, and β-actin (Cell Signaling Technology, Danvers, MA, USA; Santa Cruz Biotechnology, Dallas, TX, USA) as a loading control.

Techniques: Protein-Protein interactions, Expressing, Phospho-proteomics, Control

Journal: Journal of Translational Autoimmunity

Article Title: Exploring the immunomodulatory effects of environmental contaminants on autoimmune patients: An in vitro approach

doi: 10.1016/j.jtauto.2025.100341

Figure Lengend Snippet: Modulation of intracellular signaling pathways in HD, RA, and SLE cells exposed to environmental contaminants. Heatmap showing the relative expression and phosphorylation levels of key signaling proteins involved in immune regulation, inflammation, and cell survival (STAT1, STAT3, p38, AKT, and NFκB) and their phosphorylated forms in HD, RA, and SLE after exposure to PM, silica, and TCDD. Data are represented as z-score normalized values. Asterisks indicate statistically significant differences compared with the unstimulated control within each group (p < 0.05). AKT: protein kinase B; HD: healthy donors; NFκB: nuclear factor kappa-light-chain-enhancer of activated B cells; P: phosphorylated form; p38: p38 mitogen-activated protein kinase; PM: particulate matter; RA: rheumatoid arthritis; SLE: systemic lupus erythematosus; STAT: signal transducer and activator of transcription; TCDD: 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Article Snippet: Membranes were blocked with 5 % non-fat milk in TBS-T and incubated overnight at 4 °C with primary antibodies against phospho-AKT (Thr308), phospho-NFκB p65 (Ser536), phospho-p38 MAPK (Thr180/Tyr182), phospho-STAT1 (Tyr701), and phospho-STAT3 (Tyr705) (Cell Signaling Technology, Danvers, MA, USA; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Protein-Protein interactions, Expressing, Phospho-proteomics, Control

Journal: Frontiers in Immunology

Article Title: Second-order regulation: IFN-γ suppresses IL-17A-mediated type 3 inflammation

doi: 10.3389/fimmu.2026.1744476

Figure Lengend Snippet: IFN-γ suppresses type 3 inflammation through STAT1. (A) Schematic of experimental model used. (B) Total cell number, (C) Neutrophil, (D) TRAMs (E) Ly6C+ Monocytes, and (F) MoAMs in BALs of female and male WT and STAT1 KO mice 8 hours post treatment as enumerated using flow cytometry. Fold changes in aggregated data were calculated relative to the -IFN-γ treatment group within each sex. Unpaired t test. (G) Principal component analysis (PCA) plot of compiled BAL cellularity data from C57BL/6J WT and STAT1 KO mice based on total cell numbers and 11 distinct cell types identified by flow cytometry. (H) PCA loadings indicating the magnitude and direction of each variable’s contribution to a principal component. p value: *≤ 0.05, **≤ 0.01, ***≤ 0.001, ****≤ 0.0001. All data have n≥3 mice, 2 experiments, mean ± SEM.

Article Snippet: 6-week-old C57BL/6J (stock# 000664), IFN-γ KO (B6.129S7-Ifng tm1Ts /J, Stock# 002287), and STAT1 KO (B6.129S(Cg)-Stat1 tm1Dlv /J, Stock# 012606) mice were obtained from Jackson labs (USA) and subsequently bred in-house to ensure acclimation to local conditions.

Techniques: Flow Cytometry

Journal: Redox Biology

Article Title: Hyodeoxycholic acid attenuates atherosclerosis by antagonizing FXR and modulating the PD-1/mTORC1 signaling axis

doi: 10.1016/j.redox.2026.104096

Figure Lengend Snippet: HDCA-treated ameliorate inflammation and modulate lipid metabolism in vivo. (A) Acetyl-CoA levels decreased over time in HDCA-treated Tregs. (B – E) Quantitative real-time PCR analysis of the mRNA expression of key lipogenic genes, including (B) fatty acid synthase (FASN), (C) acetyl-CoA carboxylase (ACC), (D) SCD1, and (E) SREBP-1c, in Tregs with or without HDCA treatment. (F) Functional metabolic flux assay using 13 C-glucose tracing shows decreased enrichment of labeled β-hydroxypalmitate in the HDCA-treated group. (G) Immunoblot analysis of phosphorylated STAT1 in Tregs following HDCA exposure, and quantification statistical analysis results are presented in the bar graphs. (H) Representative IHC images show reduced IL-21 expression in atherosclerotic lesions of mice following HDCA treatment (magnification, 5 × ; scale bar, 50 μm). (I) Plasma cholesterol levels were measured by enzymatic colorimetry assay in FXR-sufficient controls and FXR KO mice ± HDCA. (J) Plasma triglyceride levels in the Vector and FXR KO groups were quantified at 0, 6, 12, and 18 h by a colorimetric assay. (K) Hemodynamic profiling assessed heart rate, systolic blood pressure (SBP), and diastolic blood pressure (DBP) for each group. (L – M) Ratios of monounsaturated to saturated fatty acids (MUFA/SFA) and polyunsaturated to saturated fatty acids (PUFA/SFA) in serum cholesterol esters (CE) and triglycerides (TG) were quantified by lipidomics in FXR-sufficient control and FXR KO mice ± HDCA. Data are presented as the mean ± SD (n = 3-5 biological replicates). ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Proteins were detected using the following antibodies: anti-CPT1a antibody (ab234111, abcam), anti-beta actin antibody (ab8226, abcam), anti-PERK antibody (ab229912, abcam), anti-ERK1+ERK2 antibody (ab184699, abcam), anti-S6K1 antibody (ab14708, abcam), anti-S6K1 (phospho T229) antibody (ab5231, abcam), Rac1/2/3 antibody (G-2) (sc-514583, Santa Cruz), anti-Calpain 1 antibody (ab108400, abcam), anti-MMP2 antibody (ab92536, abcam), anti-IL-10 antibody (ab310329, abcam), anti-ZNF671 antibody (HPA046099, Sigma-Aldrich), anti-MAPK6/ERK3 antibody (ab53277, abcam), SIAH1 recombinant rabbit monoclonal antibody (PSH01-80) (MA5-51926, Thermo Fisher), p-Stat1 antibody (pY701.4A) (sc-136229, Santa Cruz), Stat1 antibody (C-136) (sc-464, Santa Cruz), anti-FXR1 antibody (ab155124, abcam), phospho-Raptor (Ser792) polyclonal antibody (PA5-118730, Thermo Fisher), anti-PD1 antibody (ab214421, abcam), SHP-2 antibody (3752S, Cell Signaling Technology), IL-10R antibody (3F9) (sc-53654, Santa Cruz), GAPDH antibody (6C5) (sc-32233, Santa Cruz), rabbit anti-mouse IgG H&L (HRP) (ab6728, abcam), goat anti-rabbit IgG (H + L) highly cross-adsorbed secondary antibody, Alexa FluorTM Plus 488 (A32731, Thermo Fisher).

Techniques: In Vivo, Real-time Polymerase Chain Reaction, Expressing, Functional Assay, Flux Assay, Labeling, Western Blot, Clinical Proteomics, Colorimetric Assay, Plasmid Preparation, Control