Fig 1: Developmental trajectories and subgroup transitions of plasma cells exhibiting varying differentiation pathways in the pancreatic lesions of patients with type 1 autoimmune pancreatitis (AIP). A) UMAP plot displaying the annotation of distinct plasma cell subgroups identified within the dataset. B) Bar graph illustrating the relative proportions of each plasma cell subgroup present. C) Left: Pseudotime analysis revealing the differentiation trajectory among plasma cells, highlighting the progression of cell states. Right: Visualization of IGHG4 expression levels specifically in plasma cells. D) The differentiation direction of plasma cells, as determined by the Vector algorithm, is shown; the blue box indicates the proposed starting point of the differentiation process. E) Pseudotime analysis results for plasma cells, illustrating their developmental progression over time. F) Identification of DEGs and functional modules through pseudotime analysis, along with enriched GO terms (p < 0.01). G) The calculation of putative transcription factors associated with each plasma cell subgroup, derived from analysis utilizing the SENIC algorithm. H–J) Selected GO terms for CALR‐positive plasma cells, IgG4‐positive plasma cells, and PRDM1‐positive plasma cells in the type 1 AIP group compared to those in the normal pancreatic tissue or the post‐PBMC group, as determined by Gene Set Enrichment Analysis (GSEA). K,M) Left: Representative immunofluorescence images depicting CALR‐positive plasma cells and PRDM1‐positive plasma cells within the pancreatic lesions of type 1 AIP patients. Scale bars indicate 50 µm. Right: Quantification of the density of CALR‐positive plasma cells and PRDM1‐positive plasma cells in pancreatic tissues from type 1 AIP patients (n = 5) in comparison to healthy controls (n = 5) (***p < 0.001). Scale bars indicate 50 µm.
Fig 2: Silencing of circFOXP1 counteracts UC-MSCs-derived exosomes-suppressed Th2 differentiation of CD4+ T cells. A The percentage of Th2 cells were detected by flow cytometry. B The expression of IL-4, IL-5 and IL-13 in CD4+ T cells were detected by qRT-PCR. C The secretion of IL-4, IL-5 and IL-13 in CD4+ T cells were detected by ELISA assay. D The protein levels of GATA3, IL-4, EGR1 and Blimp-1 in CD4+ T cells were detected by western blot. Results were presented as mean ± SD from 3 independent experiments. A representative western blot from three independent experiments is shown. *P<0.05, **P<0.01, ***P<0.001
Fig 3: EGR1 acts as a transcriptional activator of Blimp-1. A-B The putative binding sites of EGR1 on the promoter region of Blimp-1 were predicted by JASPAR database. C The interaction between EGR1 and Blimp-1 promoter was detected by ChIP assay. Normal IgG served as a negative control. D The relative luciferase activity was measured by luciferase reporter assay. Renilla luciferase activity was used as an internal control. UC-MSCs were transfected with EGR1 overexpression plasmid. E-F The mRNA levels of EGR1 and Blimp-1 were detected by qRT-PCR. G The protein levels of EGR1 and Blimp-1 were detected by western blot. Results were presented as mean ± SD from 3 independent experiments. A representative western blot from three independent experiments is shown. **P<0.01, ***P<0.001
Fig 4: EGR1 and Blimp-1 are elevated in AR. The nasal mucosal samples were collected from healthy controls (HCs), disease controls (DCs) and AR patients (n=23). A-B The mRNA levels of EGR1 and Blimp-1 in the nasal mucosa were detected by qRT-PCR. C-D ROC curves of EGR1 and Blimp-1 in distinguishing AR patients from DCs or HCs. E-F The immunoreactivities of EGR1 and Blimps in the nasal mucosal samples were detected by IHC analysis. Scale bar, 100 μm for 100X and 25 μm for 400X. *P<0.05, **P<0.01, ***P<0.001
Fig 5: circFOXP1 triggers epigenetic repression of EGR1 to transcriptionally suppress Blimp-1 through recruiting EZH2. A The nuclear and cytoplasmic expression of circFOXP1 in CD4+ T cells was detected by qRT-PCR. The association between circFOXP1 and EZH2 in CD4+ T cells was assessed by RNA pull-down B and RIP assays C. Scramble RNA was used as a control probe for RNA pull-down assay. Normal IgG served as a negative control for RIP assay. D The subcellular co-localization of circFOXP1 and EZH2 in CD4+ T cells was detected by RNA FISH/IF staining. Scale bar, 25 μm for 400X. Red, circFOXP1; Green, EZH2; Blue, DAPI. E-F The enrichments of EZH2 and H3K27me3 in the promoter region of EGR1 were detected by ChIP assay. Normal IgG served as a negative control. G The mRNA levels of EGR1 and Blimp-1 in transfected CD4+ T cells were detected by qRT-PCR. H The protein levels of EZH2, EGR1 and Blimp-1 in transfected CD4+ T cells were detected by western blot. Results were presented as mean ± SD from 3 independent experiments. A representative western blot from three independent experiments is shown. *P<0.05, **P<0.01, ***P<0.001
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