Journal: Cell

Article Title: Self-Recognition of an Inducible Host lncRNA by RIG-I Feedback Restricts Innate Immune Response.

doi: 10.1016/j.cell.2018.03.064

Figure Lengend Snippet: Figure 2. lnc-Lsm3b Inhibits RIG-I-Mediated Signaling in RNA Virus-Infected Macrophages (A–C) ELISA of IFN-a (A), IFN-b (B), and IL-6 (C) in the supernatants of peritoneal macrophages transfected with si-ctrl or si-lnc-Lsm3b for 48 hr before VSV, SeV, or HSV-1 infection for 16 hr. (D–F) ELISA of IFN-a (D), IFN-b (E), and IL-6 (F) in supernatants of lnc-Lsm3b+/+ and lnc-Lsm3b/ cells infected with VSV for indicated hours. (G and H) IFN-b (G) and NF-kB (H) promoter activity in HEK293T cells transfected with lnc-Lsm3b and RIG-I vectors upon VSV infection. (I) VSV titers by TCID50 assay in supernatants of lnc-Lsm3b+/+ and lnc-Lsm3b/ cells infected with VSV for 12 hr and 24 hr. (J) Immunoblot analysis of RIG-I signaling pathways in lnc-Lsm3b+/+ and lnc-Lsm3b/ cells infected with VSV for indicated hours. Data are shown as mean ± SD (n = 4) or typical photographs of one representative experiment. Similar results were obtained in three independent experiments. *p < 0.05; **p < 0.01. See also Figure S3.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER poly(I:C) HMW Biotin InvivoGen Cat #tlrl-picb TRIzol reagent Invitrogen Cat #15596-018 Mouse IFN-a Miltenyi Biotec Cat #130-093-130 Mouse IFN-b R&D Systems Cat #8234-MB-010 RNA FISH probe Biosearch technologies N/A GST-RIG-I This paper N/A His-RIG-I This paper N/A GST-CTD This paper N/A GST-CARD This paper N/A His-deltaCARD This paper N/A Critical Commercial Assays NorthernMax kit Thermo Fisher Scientific Cat #AM1940 RNeasy Mini Kit QIAGEN Cat #74104 miRNeasy Serum/Plasma Spike-In Control QIAGEN Cat #219610 MAXIscript SP6/T7 Kit Ambion Cat #AM1320 Streptavidin beads Thermo Fisher Scientific Cat #11206D Mouse IFN-a ELISA kit PBL Biomedical Laboratories Cat #42120-1 Mouse IFN-b ELISA kit PBL Biomedical Laboratories Cat #42400-1 Mouse IL-6 ELISA kit R&D Systems Cat #VAL604 Dual-Luciferase assay kit Promega Cat #TM040 SMARTer RACE 50/30 kit Clontech Cat #634858 ATPase/GTPase Activity Assay Kit Sigma Cat #MAK113 Deposited Data RIP-seq data This paper GEO: GSE106254 iCLIP data This paper GEO: GSE106254 Experimental Models: Cell Lines HEK293T cells American Type Culture Collection N/A L929 cells American Type Culture Collection N/A RAW264.7 cells American Type Culture Collection N/A lnc-Lsm3b / RAW264.7 cells This paper N/A Experimental Models: Organisms/Strains lnc-Lsm3b / mice This paper N/A IFNabR / mice The Jackson Laboratory JAX stock 000819 Recombinant DNA Plasmid: pSIF vector Provided by Dr. Qian Zhang from Second Military Medical University, Shanghai, China N/A Plasmid: pGEM-3zf vector Provided by Dr. Xiaozhong Peng from Peking Union Medical College, Beijing, China N/A cDNA lnc-Lsm3b This paper N/A cDNA lnc-Lsm3b mutants This paper N/A Software and Algorithms FastQC 0.11.5 Babraham Bioinfomatics http://www.bioinformatics.babraham.ac.uk/ projects/fastqc/ Samtools 0.1.19 John Marshall and Petr Danecek http://www.htslib.org/ HISAT2 2.0.4 The Center for Computational Biology http://ccb.jhu.edu/software/hisat2/index.shtml StringTie v1.3.3b The Center for Computational Biology http://ccb.jhu.edu/software/stringtie/ (Continued on next page) e2 Cell 173, 1–14.e1–e6, May 3, 2018 Please cite this article in press as: Jiang et al., Self-Recognition of an Inducible Host lncRNA by RIG-I Feedback Restricts Innate Immune Response, Cell (2018), https://doi.org/10.1016/j.cell.2018.03.064

Techniques: Virus, Infection, Enzyme-linked Immunosorbent Assay, Transfection, Activity Assay, TCID50 Assay, Western Blot, Protein-Protein interactions

Journal: Cell

Article Title: Self-Recognition of an Inducible Host lncRNA by RIG-I Feedback Restricts Innate Immune Response.

doi: 10.1016/j.cell.2018.03.064

Figure Lengend Snippet: Figure 3. lnc-Lsm3-Deficient Mice Produce More Type I IFNs (A) Survival of 6-week-old lnc-Lsm3b+/+ and lnc-Lsm3b/ mice given VSV (1 3 108 pfu/g body weight) via tail intravenous injection (i.v.) (n = 10). Kaplan-Meier method was used to evaluate survival curves. (B) ELISA of type I IFNs (IFN-a and IFN-b) production in sera from lnc-Lsm3b+/+ and lnc-Lsm3b/ mice (n = 5) at 18 hr and 24 hr after i.v. injection with VSV (5 3 107 pfu/g). (C) VSV loads by TCID50 assay and qPCR assay of VSV RNA copies in organs and peritoneal macrophages from mice in (B) (n = 4). (D) Pathology of lnc-Lsm3b+/+ and lnc-Lsm3b/ mice in response to VSV. H&E staining of lung sections from mice in (B). Scale bar, 80 mm. (E and F) qPCR analysis of Ifn-b (E) and lnc-Lsm3b (F) expression in organs and peritoneal macrophages from mice in (B) (n = 5). Data are shown as mean ± SD or typical photographs of one representative experiment. Similar results were obtained in three independent experiments. *p < 0.05; **p < 0.01. See also Figures S3 and S4.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER poly(I:C) HMW Biotin InvivoGen Cat #tlrl-picb TRIzol reagent Invitrogen Cat #15596-018 Mouse IFN-a Miltenyi Biotec Cat #130-093-130 Mouse IFN-b R&D Systems Cat #8234-MB-010 RNA FISH probe Biosearch technologies N/A GST-RIG-I This paper N/A His-RIG-I This paper N/A GST-CTD This paper N/A GST-CARD This paper N/A His-deltaCARD This paper N/A Critical Commercial Assays NorthernMax kit Thermo Fisher Scientific Cat #AM1940 RNeasy Mini Kit QIAGEN Cat #74104 miRNeasy Serum/Plasma Spike-In Control QIAGEN Cat #219610 MAXIscript SP6/T7 Kit Ambion Cat #AM1320 Streptavidin beads Thermo Fisher Scientific Cat #11206D Mouse IFN-a ELISA kit PBL Biomedical Laboratories Cat #42120-1 Mouse IFN-b ELISA kit PBL Biomedical Laboratories Cat #42400-1 Mouse IL-6 ELISA kit R&D Systems Cat #VAL604 Dual-Luciferase assay kit Promega Cat #TM040 SMARTer RACE 50/30 kit Clontech Cat #634858 ATPase/GTPase Activity Assay Kit Sigma Cat #MAK113 Deposited Data RIP-seq data This paper GEO: GSE106254 iCLIP data This paper GEO: GSE106254 Experimental Models: Cell Lines HEK293T cells American Type Culture Collection N/A L929 cells American Type Culture Collection N/A RAW264.7 cells American Type Culture Collection N/A lnc-Lsm3b / RAW264.7 cells This paper N/A Experimental Models: Organisms/Strains lnc-Lsm3b / mice This paper N/A IFNabR / mice The Jackson Laboratory JAX stock 000819 Recombinant DNA Plasmid: pSIF vector Provided by Dr. Qian Zhang from Second Military Medical University, Shanghai, China N/A Plasmid: pGEM-3zf vector Provided by Dr. Xiaozhong Peng from Peking Union Medical College, Beijing, China N/A cDNA lnc-Lsm3b This paper N/A cDNA lnc-Lsm3b mutants This paper N/A Software and Algorithms FastQC 0.11.5 Babraham Bioinfomatics http://www.bioinformatics.babraham.ac.uk/ projects/fastqc/ Samtools 0.1.19 John Marshall and Petr Danecek http://www.htslib.org/ HISAT2 2.0.4 The Center for Computational Biology http://ccb.jhu.edu/software/hisat2/index.shtml StringTie v1.3.3b The Center for Computational Biology http://ccb.jhu.edu/software/stringtie/ (Continued on next page) e2 Cell 173, 1–14.e1–e6, May 3, 2018 Please cite this article in press as: Jiang et al., Self-Recognition of an Inducible Host lncRNA by RIG-I Feedback Restricts Innate Immune Response, Cell (2018), https://doi.org/10.1016/j.cell.2018.03.064

Techniques: Injection, Enzyme-linked Immunosorbent Assay, TCID50 Assay, Staining, Expressing

Journal: bioRxiv

Article Title: Epstein-Barr virus induced 3 attributes to TLR7-mediated splenomegaly and bicytopenia

doi: 10.1101/2024.06.10.598101

Figure Lengend Snippet: Ebi3 deficiency did not alter the development and function of plasmacytoid dendritic cells (pDCs). (A) Representative flow cytometric analysis of the percentage of pDCs (CD11c + B220 + ) and conventional dendritic cells (cDCs) (CD11c + B220 - ) in the spleen of wild-type (WT) or KO mice. Numbers in the plots indicate the percentage of cells in each gate. (B) Percentages of pDCs (CD11c + B220 + Gr-1 + ) or cDCs (CD11c + B220 - CD11b + or CD11c + B220 - CD11b - ) in the spleen of WT or KO mice. (C) Infa gene expression in ear skin sample of Vaseline- or imiquimod (IMQ)-treated WT or KO mice. Gene expression was analyzed with quantitative polymerase chain reaction (qPCR). (D) IFN-α concentration in serum of Vaseline- or IMQ-treated WT or KO mice. IFN-α concentrations were quantified using enzyme-linked immunosorbent assay (ELISA). (E, F) Bone marrow-derived pDCs were stimulated with 200 ng/ml of IMQ for 6 hours. (E) Ifna gene expression in the bone-marrow derived pDCs. Expression levels were analyzed by qPCR and normalized to that of Hprt. (F) The concentrations of IFN-α in the culture supernatants. All data are expressed as the mean + standard deviation.

Article Snippet: Concentrations of IFN-α in culture supernatants or mouse serum were measured using Mouse IFN-alpha All Subtype Quantikine ELISA Kit (MFNAS0, R&D Systems) according to the manufacturer’s protocols.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Concentration Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Expressing, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Therapeutically targeting type I interferon directly to XCR1+ dendritic cells reveals the role of cDC1s in anti-drug antibodies

doi: 10.3389/fimmu.2023.1272055

Figure Lengend Snippet: XCR1Ab-IFN fusions dramatically enhanced PK and cDC1 activation. (A) Pharmacokinetic profile comparison of mouse recombinant IFNs (IFN-alphaA/15 and IFN-beta) and Ab-IFN fusions of mouse IFN-alphaA/15 (IFN WT ). Data shown are mean. N=3 mice/group. (B) Schematic of in vivo study design. (C) Gating strategy to assess XCR1 receptor occupancy and IFN activity in cDC1s in spleen (CD8 + DCs) and tumor (CD103 + DCs). (D) Detection of XCR1 surface expression using a competing PE conjugated anti-XCR1 Ab on cDC1s in spleen and tumor. (E) XCR1 receptor occupancy was quantified by normalizing the % of XCR1 + cells detected in cDC1s in treated mice to the % of XCR1 + cells detected in vehicle control cDC1s. PD-L1 and CD86 surface expression was evaluated by flow cytometry as a readout for IFN-induced activity in (F) targeted (XCR1 + ) CD8 + or CD103 + cDC1s and (G) untargeted (XCR1-) Ly6G + neutrophils. Median fluorescent intensity (MFI) was measured. Data shown are mean +/- SD. N=4 mice/group. * P < 0.05, ** P < 0.01, *** P < 0.001, ****P < 0.0001 by 1-way ANOVA with Tukey’s multiple-comparison post hoc test (E–G) .

Article Snippet: To quantitate IFN-alpha and IFN-beta recombinant proteins (all from PBL), VeriKineTM Mouse IFN Alpha ELISA Kit (PBL) and VeriKineTM Mouse IFN Beta ELISA Kit (PBL) were used respectively.

Techniques: Activation Assay, Comparison, Recombinant, In Vivo, Activity Assay, Expressing, Control, Flow Cytometry

Journal: Inflammation

Article Title: Bifidobacterium Infantis Promotes FOSL1-Mediated Transcriptional Repression of VDR to Accelerate Autoimmune Pancreatitis Progression Through Activation of Oxidative Stress

doi: 10.1007/s10753-025-02368-4

Figure Lengend Snippet: B. Infantis induces the AIP progression in mice with mild AIP. ( A ) Inflammatory infiltration and damage in the mouse pancreas were observed using HE staining and histologic damage scoring. ( B ) Pancreatic fibrosis in mice was assessed using Picro Sirius red staining. ( C ) IHC scores of Fibronectin and α-SMA in mouse pancreatic tissues. ( D ) The proportion of activated pDCs in PMNC in mouse pancreatic tissues was detected by flow cytometry. ( E ) Activated pDCs (pDCA-1, green) in mouse pancreatic tissues were observed using immunofluorescence staining. ( F ) IFN-α, IL-33, and IL-13 levels in supernatants of mouse pancreatic tissue homogenates were assessed using ELISA. Statistical analyses were performed using one-way ANOVA, followed by Tukey’s post hoc test. Results are expressed as mean ± SEM ( n = 5). * p < 0.05.

Article Snippet: Mouse pancreatic tissues were homogenized and centrifuged to collect the supernatant, or the culture supernatant of pDCs was harvested and assayed by Mouse IFN-α ELISA Kit (NBP3-42614, Novus Biological) and Mouse IL-33 ELISA Kit (LS-F26388-1, Vector Laboratories, Newark, CA, USA) according to the instructions.

Techniques: Staining, Flow Cytometry, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Inflammation

Article Title: Bifidobacterium Infantis Promotes FOSL1-Mediated Transcriptional Repression of VDR to Accelerate Autoimmune Pancreatitis Progression Through Activation of Oxidative Stress

doi: 10.1007/s10753-025-02368-4

Figure Lengend Snippet: B. infantis activates pDCs to promote AIP in mice. ( A ) Inflammatory infiltration and damage in the mouse pancreas were observed using HE staining and histologic damage scoring. ( B ) Pancreatic fibrosis in mice was assessed using Picro Sirius red staining. ( C ) IHC scores of Fibronectin and α-SMA in mouse pancreatic tissues. ( D ) The proportion of activated pDCs in PMNC in mouse pancreatic tissues was detected by flow cytometry. ( E ) Activated pDCs (pDCA-1, green) in mouse pancreatic tissues were observed using immunofluorescence staining. ( F ) IFN-α, IL-33, and IL-13 levels in supernatants of mouse pancreatic tissue homogenates were assessed using ELISA. Statistical analyses were performed using unpaired t-tests. Results are expressed as mean ± SEM ( n = 5). * p < 0.05.

Article Snippet: Mouse pancreatic tissues were homogenized and centrifuged to collect the supernatant, or the culture supernatant of pDCs was harvested and assayed by Mouse IFN-α ELISA Kit (NBP3-42614, Novus Biological) and Mouse IL-33 ELISA Kit (LS-F26388-1, Vector Laboratories, Newark, CA, USA) according to the instructions.

Techniques: Staining, Flow Cytometry, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Inflammation

Article Title: Bifidobacterium Infantis Promotes FOSL1-Mediated Transcriptional Repression of VDR to Accelerate Autoimmune Pancreatitis Progression Through Activation of Oxidative Stress

doi: 10.1007/s10753-025-02368-4

Figure Lengend Snippet: B. infantis enhances poly (I: C)-stimulated activation of pDCs in vitro. ( A ) Detection of IFN-α and IL-33 secreted by pDCs isolated from mice with no treatment (control), high dose of poly (I: C) (severe inflammation), and low dose of poly (I: C) without further treatment (mild inflammation), treated with medium (mild inflammation + medium), as well as B. infantis (mild inflammation + B. infantis ). ( B ) Immunofluorescence labeling of pDCs surface activation marker pDCA-1 (green). ( C ) Activated pDCs (Siglec-H + ) in total pDCs were analyzed using flow cytometry. ( D ) The interaction between CFSE-labeled B. infantis (green) and pDCA-1 APC antibody-stained pDCs (red, cell membrane) was observed using fluorescence microscopy. ( E ) The colocalization of LysoTracker-labeled lysosomes (red) and CFSE-labeled B. infantis (green) within pDCs was observed using fluorescence microscopy. ( F ) The schematic diagram illustrates the direct interaction pathway between B. infantis and pDCs (solid line) and the hypothesized indirect pathway (dashed line). Statistical analyses were performed using one-way ANOVA. The results shown are representative of data derived from 3 independent experiments and are expressed as mean ± SEM. * p < 0.05.

Article Snippet: Mouse pancreatic tissues were homogenized and centrifuged to collect the supernatant, or the culture supernatant of pDCs was harvested and assayed by Mouse IFN-α ELISA Kit (NBP3-42614, Novus Biological) and Mouse IL-33 ELISA Kit (LS-F26388-1, Vector Laboratories, Newark, CA, USA) according to the instructions.

Techniques: Activation Assay, In Vitro, Isolation, Control, Immunofluorescence, Labeling, Marker, Flow Cytometry, Staining, Membrane, Fluorescence, Microscopy, Derivative Assay

Journal: Inflammation

Article Title: Bifidobacterium Infantis Promotes FOSL1-Mediated Transcriptional Repression of VDR to Accelerate Autoimmune Pancreatitis Progression Through Activation of Oxidative Stress

doi: 10.1007/s10753-025-02368-4

Figure Lengend Snippet: Knockdown of FOSL1 inhibits AIP in mice administered with B. infantis . ( A ) The volcano map of DEGs screened out from pancreatic-infiltrating pDCs in mice from the Mild AIP + B. infantis or the Mild AIP + Vector group. ( B ) Acquisition of intersecting genes of the top 50 DEGs from transcriptome sequencing (ranked by adjusted p-value ) and the TF list (Mus musculus) by Jvenn. ( C ) FOSL1 (red) expression within activated pDCs (marked by pDCA-1, green) in mouse pancreatic tissues was observed using dual-labeling immunofluorescence. ( D ) IHC scores of FOSL1 in mouse pancreatic tissues. ( E ) Inflammatory infiltration and damage in the mouse pancreas were observed using HE staining and histologic damage scoring. ( F ) Pancreatic fibrosis in mice was assessed using Picro Sirius red staining. ( G ) IHC scores of Fibronectin and α-SMA in mouse pancreatic tissues. ( H ) The proportion of activated pDCs in PMNC in mouse pancreatic tissues was detected by flow cytometry. ( I ) Activated pDCs (pDCA-1, green) in mouse pancreatic tissues were observed using immunofluorescence staining. ( J ) IFN-α, IL-33, and IL-13 levels in supernatants of mouse pancreatic tissue homogenates were assessed using ELISA. For two sets of data that do not follow a normal distribution, the Mann-Whitney U test was used to compare the two groups. Comparisons between datasets conforming to normal distribution were conducted using one-way ANOVA, followed by Tukey’s post hoc test or unpaired t-tests. Results are expressed as mean ± SEM ( n = 5). * p < 0.05.

Article Snippet: Mouse pancreatic tissues were homogenized and centrifuged to collect the supernatant, or the culture supernatant of pDCs was harvested and assayed by Mouse IFN-α ELISA Kit (NBP3-42614, Novus Biological) and Mouse IL-33 ELISA Kit (LS-F26388-1, Vector Laboratories, Newark, CA, USA) according to the instructions.

Techniques: Knockdown, Plasmid Preparation, Sequencing, Expressing, Labeling, Immunofluorescence, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Inflammation

Article Title: Bifidobacterium Infantis Promotes FOSL1-Mediated Transcriptional Repression of VDR to Accelerate Autoimmune Pancreatitis Progression Through Activation of Oxidative Stress

doi: 10.1007/s10753-025-02368-4

Figure Lengend Snippet: Activation of pDCs induced by B. infantis is reversed by knockdown of FOSL1. ( A ) The protein expression of FOSL1 in pDCs isolated from mice with no treatment (control), a high dose of poly (I: C) (severe inflammation), and a low dose of poly (I: C) without further treatment (mild inflammation), treated with medium (mild inflammation + medium), as well as B. infantis (mild inflammation + B. infantis ) by western blot analysis. ( B ) The protein expression of FOSL1 in mildly inflamed pDCs treated with B. infantis and sh-FOSL1 or sh-Scramble was analyzed by western blot analysis. ( C ) IFN-α and IL-33 levels in the supernatant of pDCs were examined using ELISA. ( D ) FOSL1 expression in isolated pDCs (marked by pDCA-1) was observed using dual-labeling immunofluorescence. ( E ) Activated pDCs (Siglec-H + ) in total pDCs were analyzed using flow cytometry. Statistical analyses were performed using one-way ANOVA, followed by Tukey’s post hoc test or unpaired t-tests. The results shown are representative of data derived from 3 independent experiments and are expressed as mean ± SEM. * p < 0.05.

Article Snippet: Mouse pancreatic tissues were homogenized and centrifuged to collect the supernatant, or the culture supernatant of pDCs was harvested and assayed by Mouse IFN-α ELISA Kit (NBP3-42614, Novus Biological) and Mouse IL-33 ELISA Kit (LS-F26388-1, Vector Laboratories, Newark, CA, USA) according to the instructions.

Techniques: Activation Assay, Knockdown, Expressing, Isolation, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Labeling, Immunofluorescence, Flow Cytometry, Derivative Assay

Journal: Inflammation

Article Title: Bifidobacterium Infantis Promotes FOSL1-Mediated Transcriptional Repression of VDR to Accelerate Autoimmune Pancreatitis Progression Through Activation of Oxidative Stress

doi: 10.1007/s10753-025-02368-4

Figure Lengend Snippet: B. infantis promotes oxidative stress in pDCs and exacerbates AIP by inhibiting VDR transcription. ( A ) Inflammatory infiltration and damage in the mouse pancreas were observed using HE staining and histologic damage scoring. ( B ) Pancreatic fibrosis in mice was assessed using Picro Sirius red staining. ( C ) IHC scores of Fibronectin, α-SMA, and VDR in mouse pancreatic tissues. ( D ) The proportion of activated pDCs in PMNC in mouse pancreatic tissues was detected by flow cytometry. ( E ) VDR (red) expression in isolated pDCs (marked by pDCA-1, green) was observed using dual-labeling immunofluorescence. ( F ) IFN-α, IL-33, and IL-13 levels in supernatants of mouse pancreatic tissue homogenates were assessed using ELISA. ( G ) MDA and SOD concentrations in the supernatant of mouse pancreatic tissue homogenate. ( H ) Analysis of H 2 O 2 and GSH levels in mouse pancreatic tissue homogenate supernatant using ELISA kits. Statistical analyses were performed using unpaired t-tests. Results are expressed as mean ± SEM ( n = 5). * p < 0.05.

Article Snippet: Mouse pancreatic tissues were homogenized and centrifuged to collect the supernatant, or the culture supernatant of pDCs was harvested and assayed by Mouse IFN-α ELISA Kit (NBP3-42614, Novus Biological) and Mouse IL-33 ELISA Kit (LS-F26388-1, Vector Laboratories, Newark, CA, USA) according to the instructions.

Techniques: Staining, Flow Cytometry, Expressing, Isolation, Labeling, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Inflammation

Article Title: Bifidobacterium Infantis Promotes FOSL1-Mediated Transcriptional Repression of VDR to Accelerate Autoimmune Pancreatitis Progression Through Activation of Oxidative Stress

doi: 10.1007/s10753-025-02368-4

Figure Lengend Snippet: Knockdown of VDR expedites oxidative stress in pDCs in the presence of FOSL1 knockdown. ( A ) The content of IFN-α and IL-33 secreted by pDCs transduced with sh-FOSL1 + sh-Scramble or sh-VDR was examined using ELISA. ( B ) VDR expression in isolated pDCs (marked by pDCA-1) was observed using dual-labeling immunofluorescence. ( C ) Activated pDCs (Siglec-H + ) in total pDCs were analyzed using flow cytometry. ( D ) Analysis of ROS content within mouse pDCs by DCFH-DA probe. Statistical analyses were performed using unpaired t-tests. The results shown are representative of data derived from 3 independent experiments and are expressed as mean ± SEM. * p < 0.05.

Article Snippet: Mouse pancreatic tissues were homogenized and centrifuged to collect the supernatant, or the culture supernatant of pDCs was harvested and assayed by Mouse IFN-α ELISA Kit (NBP3-42614, Novus Biological) and Mouse IL-33 ELISA Kit (LS-F26388-1, Vector Laboratories, Newark, CA, USA) according to the instructions.

Techniques: Knockdown, Transduction, Enzyme-linked Immunosorbent Assay, Expressing, Isolation, Labeling, Immunofluorescence, Flow Cytometry, Derivative Assay

Journal: Inflammation

Article Title: Bifidobacterium Infantis Promotes FOSL1-Mediated Transcriptional Repression of VDR to Accelerate Autoimmune Pancreatitis Progression Through Activation of Oxidative Stress

doi: 10.1007/s10753-025-02368-4

Figure Lengend Snippet: Molecular mechanism. B. infantis colonization mediates oxidative stress in pDCs through activation of FOSL1 and transcriptional repression of VDR, which promotes pDCs activation and secretion of IFN-α and IL-33, accelerating AIP progression.

Article Snippet: Mouse pancreatic tissues were homogenized and centrifuged to collect the supernatant, or the culture supernatant of pDCs was harvested and assayed by Mouse IFN-α ELISA Kit (NBP3-42614, Novus Biological) and Mouse IL-33 ELISA Kit (LS-F26388-1, Vector Laboratories, Newark, CA, USA) according to the instructions.

Techniques: Activation Assay