e el r0545  (Elabscience Biotechnology)

Journal: The Journal of Biological Chemistry

Article Title: Elongator is required for pattern recognition receptor and type I interferon signaling in macrophages

doi: 10.1016/j.jbc.2025.110916

Figure Lengend Snippet: Differential protein expression in Elp3 −/− BMDMs relative to WT cells . Volcano plots showing differentially expressed proteins in unstimulated ( A ), 6 h LPS-treated ( B ) or 12 h LPS-treated ( C ) Elp3 −/− lysates versus lysates from WT cells. To capture a broad range of biological processes potentially influenced by loss of ELP3 an enrichment cut-off of a –log10 ( p -value) of 1.3 was considered significant. Proteins associated with IFN signalling by gene ontology biological process terms are labelled as red dots . IFN, interferon; LPS, lipopolysaccharide.

Article Snippet: ELISA plates were coated with a monoclonal rat anti-mouse IFNβ capture antibody (Santa Cruz, Cat#SC-57201) in carbonate buffer overnight at 4 °C.

Techniques: Expressing

Journal: The Journal of Biological Chemistry

Article Title: Elongator is required for pattern recognition receptor and type I interferon signaling in macrophages

doi: 10.1016/j.jbc.2025.110916

Figure Lengend Snippet: Impaired LPS-IFN I signalling axis in Elp3 −/− BMDMs . A , schematic of LPS- IFN-I signalling axis. Figure created in BioRender ( B – K ) Label free quantification intensity values detected by mass spectrometry in WT and Elp3 −/− cells stimulated for 0, 6 or 12 h with LPS for peptides from transcription factor proteins IRF3 ( B ), IRF5 ( C ), IRF7 ( D ), IRF9 ( E ), RELA ( F ), NFKB1 ( G ), NFKB2 ( H ), STAT1 ( I ), STAT3 ( J ) and STAT6 ( K ). ND, not detected. Data are mean ± SD for quadruplicate (or triplicate for WT 0 h sample) measurements. ∗ p < 0.05 compared to WT, based on Mann-Whitney test. L – N , WT and Elp3 −/− cells were stimulated with LPS (100 ng/ml) for 3, 6 and 24 h. Ifnb ( L ), Irf7 ( M ) and Ifna ( N ) mRNA were assayed by qRT-PCR, expressed relative to the untreated WT control and normalized to the housekeeping gene β-actin. Data are mean ± SD of three independent experiments. Data significance was tested with a 2-way ANOVA using Šídák's multiple comparisons test. ∗ p < 0.05 and ∗∗ p < 0.0001 compared to WT. O , cells were stimulated with LPS for the indicated times and supernatants assayed by ELISA for IFN-β. Data are mean ± SD for three independent experiments, each performed on three WT or Elp3 −/− clones in triplicate. ∗∗∗ p < 0.001. LPS, lipopolysaccharide.

Article Snippet: ELISA plates were coated with a monoclonal rat anti-mouse IFNβ capture antibody (Santa Cruz, Cat#SC-57201) in carbonate buffer overnight at 4 °C.

Techniques: Quantitative Proteomics, Mass Spectrometry, MANN-WHITNEY, Quantitative RT-PCR, Control, Enzyme-linked Immunosorbent Assay, Clone Assay

Journal: The Journal of Biological Chemistry

Article Title: Elongator is required for pattern recognition receptor and type I interferon signaling in macrophages

doi: 10.1016/j.jbc.2025.110916

Figure Lengend Snippet: ELP3 is required for IFN I and TYK 2-dependent signaling . A , WT and Elp3 −/− BMDMs were stimulated with 1000 U/ml IFNβ for the indicated times. Cells were harvested and immunoblotted for phosphorylated STAT1, STAT1 and β-actin. Representative of three independent experiments. B – D , WT and Elp3 −/− BMDMs were stimulated with 1000 U/ml IFNβ for the indicated times. RNA was isolated and mRNA expression of Irf7 ( B ), Isg15 ( C ), and Stat1 ( D ) were assayed by qRT-PCR, expressed relative to the untreated WT control and normalized to β-actin. E , WT and Elp3 −/− iBMDMs were stimulated with either LPS (100 ng/ml), IFNβ (1000 U/ml) or IFNγ (25 ng/ml) for 90 min. Cells were harvested and expression of the indicated proteins was assessed by immunoblot. Representative of 3 independent experiments. F , WT and Elp3 −/− iBMDMs were stimulated with either LPS (100 ng/ml), IFNβ (1000 U/ml) or IFNγ (25 ng/ml) for 3 h. Irf1 gene expression was assessed by qRT-PCR. mRNA levels are represented relative to β-actin, with each WT sample set to 100% and Elp3 −/− mRNA levels expressed as a percentage of their WT stimulated counterpart. G , WT and Elp3 −/− iBMDMs were stimulated with IL-10 (10 ng/ml) for 3 h. Socs3 mRNA was assayed by qRT-PCR, expressed relative to the untreated WT control and normalized to the housekeeping gene β-actin. H , WT and Elp3 −/− iBMDMs were stimulated with IL-4 (10 ng/ml) for 3 h. Arg1 mRNA was assayed by qRT-PCR, expressed relative to the untreated WT control and normalized to the housekeeping gene β-actin. For ( B – D and F – H ), data are mean ± SD of 3 independent experiments, each performed in triplicate. Data significance was tested with a 2-way ANOVA using Šídák's multiple comparisons test. ∗ p < 0.05 and ∗∗ p < 0.01 and ∗∗∗ p < 0.001 compared to WT. iBMDM, immortalised bone marrow–derived macrophage; LPS, lipopolysaccharide.

Article Snippet: ELISA plates were coated with a monoclonal rat anti-mouse IFNβ capture antibody (Santa Cruz, Cat#SC-57201) in carbonate buffer overnight at 4 °C.

Techniques: Isolation, Expressing, Quantitative RT-PCR, Control, Western Blot, Gene Expression, Derivative Assay

Journal: Molecular Medicine Reports

Article Title: Treatment with human placental extracts inhibits allergic rhinitis by modulating AMPK/SHP1/SHP2/STING signaling

doi: 10.3892/mmr.2025.13548

Figure Lengend Snippet: HPE can alleviate allergic rhinitis damage, with AMPK serving a notable role. (A) Hematoxylin and eosin staining of nasal mucosal tissues. (B) ELISA was used to detect IL-1β, IFNβ, IgE, IgG1 and IgG2 levels in the serum of rats. (C) Immunofluorescence was used to detect the expression levels of CD68 in the rats. **P<0.01, *P<0.05. HPE, human placental extracts; IFN, interferon; Ig, immunoglobulin. **P<0.01, *P<0.05.

Article Snippet: The ELISA kit includes Interleukin-1β (IL-1β) (PI303, Beyotime), Interferon-β (IFNβ) (E-EL-R0545, Elabscience), Interferon-α (IFNα) (E-EL-R3036, Elabscience), malondialdehyde (MDA) (S0131S, Beyotime), superoxide dismutase (SOD) (E-EL-R1424, Elabscience), Immunoglobulin E (IgE) (E-EL-R0517, Elabscience), Immunoglobulin G1 (IgG1) (88-50500-88, Thermo Fisher Scientific), Immunoglobulin G2a (IgG2a) (E-EL-R2415, Elabscience) and glutathione (GSH) (E-EL-0026, Elabscience).

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Expressing

Journal: Molecular Medicine Reports

Article Title: Treatment with human placental extracts inhibits allergic rhinitis by modulating AMPK/SHP1/SHP2/STING signaling

doi: 10.3892/mmr.2025.13548

Figure Lengend Snippet: HPE alleviates the inflammatory response. (A) Immunofluorescence staining was used to detect extracted alveolar macrophages. (B) Western blot analysis was conducted to detect the protein expression levels of NOX4, p-AMPK/t-AMPK, p-SHP1/t-SHP1 and p-SHP2/t-SHP2 in macrophages. (C) Western blot analysis was conducted to detect the protein expression levels of p-STING/t-STING, p-TBK1/t-TBK1, NLRP3 and IFNβ in macrophages. ELISA was used to measure the levels of (D) MDA, SOD and GSH, and (E) IL-1β, IFNβ and IFNα in macrophage supernatants. **P<0.01, *P<0.05, ns P>0.05. GSH, glutathione; HPE, human placental extracts; IFN, interferon; LPS, lipopolysaccharide; MDA, malondialdehyde; NC, negative control; NOX4, NADPH oxidase 4; p-, phosphorylated; SHP, Src homology 2-containing phosphatase; SOD, superoxide dismutase; t-t, total.

Article Snippet: The ELISA kit includes Interleukin-1β (IL-1β) (PI303, Beyotime), Interferon-β (IFNβ) (E-EL-R0545, Elabscience), Interferon-α (IFNα) (E-EL-R3036, Elabscience), malondialdehyde (MDA) (S0131S, Beyotime), superoxide dismutase (SOD) (E-EL-R1424, Elabscience), Immunoglobulin E (IgE) (E-EL-R0517, Elabscience), Immunoglobulin G1 (IgG1) (88-50500-88, Thermo Fisher Scientific), Immunoglobulin G2a (IgG2a) (E-EL-R2415, Elabscience) and glutathione (GSH) (E-EL-0026, Elabscience).

Techniques: Immunofluorescence, Staining, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Negative Control

Journal: Molecular Medicine Reports

Article Title: Treatment with human placental extracts inhibits allergic rhinitis by modulating AMPK/SHP1/SHP2/STING signaling

doi: 10.3892/mmr.2025.13548

Figure Lengend Snippet: HPE reduces cell migration. (A) Transwell experiment assessed the migration of macrophages. (B) Statistical analysis of the migration assay. **P<0.01, *P<0.05. ns; not significant (P>0.05); HPE, human placental extracts; IFN, interferon; LPS, lipopolysaccharide; NC, negative control.

Article Snippet: The ELISA kit includes Interleukin-1β (IL-1β) (PI303, Beyotime), Interferon-β (IFNβ) (E-EL-R0545, Elabscience), Interferon-α (IFNα) (E-EL-R3036, Elabscience), malondialdehyde (MDA) (S0131S, Beyotime), superoxide dismutase (SOD) (E-EL-R1424, Elabscience), Immunoglobulin E (IgE) (E-EL-R0517, Elabscience), Immunoglobulin G1 (IgG1) (88-50500-88, Thermo Fisher Scientific), Immunoglobulin G2a (IgG2a) (E-EL-R2415, Elabscience) and glutathione (GSH) (E-EL-0026, Elabscience).

Techniques: Migration, Negative Control

Journal: Molecular Medicine Reports

Article Title: Treatment with human placental extracts inhibits allergic rhinitis by modulating AMPK/SHP1/SHP2/STING signaling

doi: 10.3892/mmr.2025.13548

Figure Lengend Snippet: HPE inhibits allergic rhinitis by modulating the AMPK/SHP1/SHP2/STING signaling pathway. HPE promotes the NF-κB signaling pathway through activation of AMPK, and NF-κB directly activates SHP2 on the one hand, and SHP1 on the other hand via C/EBPα/PU.1. SHP1 inhibits STING, and SHP2 inhibits TBK1, thereby inhibiting the STING/TBK1 signaling pathway, which mediates inflammatory responses and promotes mitochondrial via IRF3 Oxidative Stress. Mitochondrial oxidative stress upregulates the expression of cGAMP and P53: cGAMP binds to STING and stabilizes its dimers and oligomers; P53 inhibits SHP1 and SHP2, and promotes the expression of both SYK and EGFR. STING interacts with EGFR, leading to the autophosphorylation of EGFR and activation of SYK, which then phosphorylates STING and EGFR at Y240 and Y245 sites, respectively. SYK and EGFR phosphorylate STING at the Y240 and Y245 sites, respectively, promoting its translocation to ERGIC for signaling. cGAMP, cyclic GMP-AMP; EGFR, epidermal growth factor; HPE, human placental extracts; IFN, interferon; IRF3, IFN regulatory factor 3; OVA, ovalbumin; SHP, Src homology 2-containing phosphatase; SYK, spleen tyrosine kinase.

Article Snippet: The ELISA kit includes Interleukin-1β (IL-1β) (PI303, Beyotime), Interferon-β (IFNβ) (E-EL-R0545, Elabscience), Interferon-α (IFNα) (E-EL-R3036, Elabscience), malondialdehyde (MDA) (S0131S, Beyotime), superoxide dismutase (SOD) (E-EL-R1424, Elabscience), Immunoglobulin E (IgE) (E-EL-R0517, Elabscience), Immunoglobulin G1 (IgG1) (88-50500-88, Thermo Fisher Scientific), Immunoglobulin G2a (IgG2a) (E-EL-R2415, Elabscience) and glutathione (GSH) (E-EL-0026, Elabscience).

Techniques: Activation Assay, Expressing, Translocation Assay