irf4  (Cell Signaling Technology Inc)

Journal: Bioactive Materials

Article Title: Immunomodulatory supramolecular hydrogel for rheumatoid arthritis management via adenosine A2A receptor-mediated macrophage remodeling

doi: 10.1016/j.bioactmat.2025.11.031

Figure Lengend Snippet: Macrophage remodeling via MTX@PNSH therapy elicits the reestablishment of immune homeostasis at the transcriptional level. (A) qRT-PCR analysis of relative mRNA of Irf3 with different treatments. (B) qRT-PCR analysis of relative mRNA of Irf4 with different treatments. (C) qRT-PCR analysis of relative mRNA of Irf5 with different treatments. (D) qRT-PCR analysis of relative mRNA of Irf8 with different treatments. (E) Heatmap analysis was performed to compare the gene expression of Irf3 , Irf4 , Irf5 , and Irf8 across different treatment groups. (F) Western blotting analysis of IRF4, and IRF8 levels in macrophages after 24 h of different treatment. (G) Quantitative analysis of IRF4 protein expression. (H) Quantitative analysis of IRF8 protein expression. Data are presented as mean ± SD, n = 3. Statistical significance was determined using one-way ANOVA, followed by Dunnett's post hoc test for comparisons between groups.

Article Snippet: Subsequently, 10 mg of protein was loaded into 10 % SDS-polyacrylamide gel electrophoresis for detection of adenosine A2AR (sc-32261, Santa Cruz), IRF4 (#62834S, Cell Signaling Technology, MA, USA), IRF8 (#5628T, Cell Signaling Technology, MA, USA) and GAPDH (TA-08, ZSGB-BIO, Beijing, China).

Techniques: Quantitative RT-PCR, Gene Expression, Western Blot, Expressing

Journal: bioRxiv

Article Title: A Novel Monocyte-derived Antigen Presenting Cell-T regulatory Cell Axis Contributes to Skin Wound healing and is Impaired in Diabetic Mice

doi: 10.64898/2026.03.04.709590

Figure Lengend Snippet: ( A ) Irf4 expression highly enriched in APC cluster compared to other clusters. ( B ) Schematic for crossbreeding B6.129S1- Irf4 tm1Rdf /J ( Irf4 fl/fl ) with C57BL/6-Tg( Csf1r -cre)1Mnz/J mice to generate Csf1r Cre Irf4 fl/fl mice. ( C ) Left: Cre-mediated recombination in Csf1r Cre Irf4 fl/fl mice confirmed by flow cytometry assessment of Green Fluorescent Protein (GFP) expression. Right: Flow cytometry analysis of IRF4+ cells indicated ablation of Irf4 in GFP+ cells compared to GFP-cells in skin wounds of Cfs1r Cre Irf4 fl/fl mice (n=6/group from two independent experiments). Percentages of APC ( D ), Ly6C hi Mo/Mφ ( E ), Ly6C int/- Mφ ( F ), and neutrophils ( G ) in wounds from Csf1 Cre Irf4 fl/fl and littermate control Irf4 fl/fl mice on day 3 and 6 post-injury ( Irf4 fl/fl : D3, n=11; D6, n=14; Csf1r Cre Irf4 fl/fl : n=16/group from three independent experiments). ( H ) Csf1r Cre Irf4 fl/fl mice showed delayed wound closure compared to Irf4 fl/fl mice ( Irf4 fl/fl : n=18/group; Csf1r Cre Irf4 fl/fl : n=24/group from three independent experiments). ( I ) Representative images of hematoxylin-eosin–stained (H&E) stained cryosections on day 6 post-injury. ep, epithelium; gt, granulation tissue. Arrows indicate ends of epithelial tongues. Arrowheads indicate the first hair follicle as the original wound, scale bar = 0.5mm. Re-epithelization ( J ) and granulation ( K ) evaluated in H&E stained cryosections of wounds. ( L ) Collagen deposition evaluated by trichrome staining (n=8/group from two independent experiments). Data are mean ± SEM; *P<0.05, **P<0.01, ***P<0.001, and ****P < 0.0001 between two genotypes. β: significant main effect of genotype.

Article Snippet: C57BL6/J (Non-diabetic, ND), BKS.Cg- Dock7 m +/+ Lepr db /J (Diabetic, DB), B6.129S4- Ccr2 tm1Ifc /J (CCR2 knockout), B6.SJL- Ptprc a Pepc b /BoyJ (CD45.1), B6.129S1- Irf4 tm1Rdf /J ( Irf4 fl/fl ), and C57BL/6-Tg(Csf1r-cre)1Mnz/J mice were purchased from the Jackson Laboratory (Bar Harbor, ME) and housed for at least two weeks before experiments.

Techniques: Expressing, Flow Cytometry, Control, Staining

Journal: bioRxiv

Article Title: A Novel Monocyte-derived Antigen Presenting Cell-T regulatory Cell Axis Contributes to Skin Wound healing and is Impaired in Diabetic Mice

doi: 10.64898/2026.03.04.709590

Figure Lengend Snippet: ( A ) Gating strategy for identifying Foxp3+Tregs (live CD11b+Ly6G-CD4+Foxp3+) and CD25 hi Tregs (live CD11b+Ly6G-CD4+Foxp3+CD25 hi ). Percentages of wound CD4+Foxp3+Tregs ( B ) and CD25 hi Tregs ( C ) evaluated on day 3 and 6 post-injury ( Irf4 fl/fl : D3, n=5; D6, n=6; Csf1r Cre Irf4 fl/fl : n=8/group from two independent experiments). ( D ) Schematic of adoptive transfer of bone marrow (BM) Mo from CD45.1 congenic mice to skin wounds in Csf1r Cre Irf4 fl/fl mice. ( E ) CD45.1+BM Mo-treated wounds showed accelerated wound closure compared to DPBS-treated control Csf1r Cre Irf4 fl/fl mice. Percentages of APCs ( F ), CD4+Foxp3+Tregs ( G ), and activated CD25 hi Tregs ( H ) were significantly elevated in wounds in response to CD45.1+BM Mo treatment compared to the DPBS-treatment on day 6 post-injury. CD45.1+BM Mo-treated wounds: n=8/group; DPBS-treated controls: n=6/group from two independent experiments; Data are mean ± SEM; *P<0.05, **P<0.01, ***P<0.001, and ****P < 0.0001 between two genotypes.

Article Snippet: C57BL6/J (Non-diabetic, ND), BKS.Cg- Dock7 m +/+ Lepr db /J (Diabetic, DB), B6.129S4- Ccr2 tm1Ifc /J (CCR2 knockout), B6.SJL- Ptprc a Pepc b /BoyJ (CD45.1), B6.129S1- Irf4 tm1Rdf /J ( Irf4 fl/fl ), and C57BL/6-Tg(Csf1r-cre)1Mnz/J mice were purchased from the Jackson Laboratory (Bar Harbor, ME) and housed for at least two weeks before experiments.

Techniques: Adoptive Transfer Assay, Control

Journal: bioRxiv

Article Title: A Novel Monocyte-derived Antigen Presenting Cell-T regulatory Cell Axis Contributes to Skin Wound healing and is Impaired in Diabetic Mice

doi: 10.64898/2026.03.04.709590

Figure Lengend Snippet: ( A ) Protein expression of IL27 in APC and ( B ) IL10 in CD4+CD25 hi T cells (n=6/group from two independent experiments). Due to assay limitations, Foxp3 could not be included in this panel. Wound levels of ( C ) IL27 ( Irf4 fl/fl : D3, n=5; n=7 for other groups from two independent experiments and ( D ) IL10 (n=7/group from two independent experiments) measured in wound homogenates on day 3 and 6 post-injury. ( E ) Schematic of local IL27 treatment in skin wounds of Csf1r Cre Irf4 fl/fl mice. ( F ) IL27-treated wounds showed accelerated wound closure compared to controls. IL27 treatment increased the percentage of Tregs ( G ), with a trend toward enhanced accumulation of activated CD25 hi Tregs in wounds ( H ). N=8 per group; Data are mean ± SEM; *P<0.05, **P<0.01, ***P<0.001, and ****P < 0.0001 between two genotypes; β: significant main effect of genotype.

Article Snippet: C57BL6/J (Non-diabetic, ND), BKS.Cg- Dock7 m +/+ Lepr db /J (Diabetic, DB), B6.129S4- Ccr2 tm1Ifc /J (CCR2 knockout), B6.SJL- Ptprc a Pepc b /BoyJ (CD45.1), B6.129S1- Irf4 tm1Rdf /J ( Irf4 fl/fl ), and C57BL/6-Tg(Csf1r-cre)1Mnz/J mice were purchased from the Jackson Laboratory (Bar Harbor, ME) and housed for at least two weeks before experiments.

Techniques: Expressing