early growth response protein 1  (Santa Cruz Biotechnology)

Journal: Nature Communications

Article Title: DPEP2 suppresses hyperinflammation via metabolic reprogramming of macrophages in sepsis

doi: 10.1038/s41467-026-70466-4

Figure Lengend Snippet: a ChIP-Seq profiles from ENCODE database identified a potential binding site for early growth response 1 (EGR1) in DPEP2 promoter region. b EGR1 is an inflammation-inducible transcriptional repressor responsible for suppressing DPEP2 expression in macrophages during sepsis. This image was created in BioRender. Qiu, Z. (2026) https://biorender.com/t7mfnvo . c Levels of Egr1 were determined after LPS stimulation for 0 or 12 h in RAW264.7 cells ( n = 3 biological replicates). d The expression of Dpep2 of RAW264.7 cells pre-treated with an inhibitor targeting EGR1 for 0 or 1 h, followed by LPS stimulation for 0 or 12 h, were determined ( n = 3 biological replicates). e Levels of Dpep2 were determined after LPS stimulation for 0 or 12 h in RAW264.7 cells with Egr1 deficiency (sh Egr1 ) vs control RAW264.7 cells (shNT) ( n = 3 biological replicates). f Cysteine leukotrienes and prostaglandin E2 (PGE2) synthesis pathways. g – j Levels of leukotriene D4 (LTD4) and leukotriene E4 (LTE4) secreted by RAW264.7 cells with Dpep2 deficiency (sh Dpep2 RAW264.7 cells) vs control RAW264.7 cells (shNT RAW264.7 cells) ( g , h ), and RAW264.7 cells overexpressing Dpep2 (DPEP2) vs control vector (Vector) cells ( i , j ) ( n = 3 biological replicates). k RAW264.7 cells were treated with 1 μg/mL LPS + 1 μM LTD4, or 1 μg/mL LPS + 1 μM LTE4, or 1 μg/mL LPS + DMSO for 12 h. Then, levels of Il1b , Il6, and Tnf were determined ( n = 3 biological replicates). l Flag-tagged WT DPEP2 plasmid and the enzymatic mutant targeting E190 (E190Q) were constructed. m RAW264.7 cells overexpressing WT DPEP2 (WT), enzymatic mutant DPEP2 and relatively control (Vector) were constructed. Protein levels of Flag in these cells were examined ( n = 3 or 6 biological replicates). n, o ELISA was used to measure LTD4 and LTE4 levels secreted by Vector, WT, and E190Q cells ( n = 3 biological replicates). p–r Il1b , Il6, and Tnf levels in Vector, WT, and E190Q cells were measured by quantitative real-time PCR after 12h of LPS stimulation (n=3 biological replicates). Data represent the mean ± standard deviation. ns, not significant. Two-sided unpaired Student’s t test was used in ( c ), one way ANOVA followed by the Bonferroni multiple comparisons was used in ( d, e, k, m–r ) and two-way repeated measures ANOVA with Tukey’s multiple comparisons test was used in ( g–j ).

Article Snippet: RAW264.7 cells were treated with a EGR1 inhibitor (5 μM EGR-1-IN-1 (IT25); HY-163731, MedChemExpress), CysLTR1 inhibitor (5 μM Zafrilukast; HY-17492, MedChemExpress), CysLTR2 inhibitor (1 μM HAMI3379; HY-112248A MedChemExpress), NF-κB inhibitor (30 μM JSH-23; HY-13982, MedChemExpress), ERK inhibitor (1 μM U0126; HY-12031A, MedChemExpress), JNK inhibitor (25 μM SP600125; HY-12041, MedChemExpress), p38 MAPK inhibitor (30 μM SB203580; HY-10256, MedChemExpress), or AKT inhibitor (30 μM Ly294002; HY-10108, MedChemExpress), for 1 h and stimulated with LPS (1 μg/mL) for 12 h.

Techniques: ChIP-sequencing, Binding Assay, Expressing, Control, Plasmid Preparation, Mutagenesis, Construct, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Communications Biology

Article Title: Early growth response 1 promotes RNA polymerase I-directed transcription and cancer growth by activating RRN3 expression

doi: 10.1038/s42003-025-09485-8

Figure Lengend Snippet: A – C Immunofluorescence (IF) colocalization analysis of EGR1 and Pol Ⅰ transcription machinery factors, including UBF ( A ), RRN3 ( B ) and RPA40 ( C ); IF assays were performed with HeLa cells. Scale bars in the images represent 5 μm. D – F Analysis of immunofluorescence colocalization between EGR1 and components of the Pol Ⅰ transcription machinery, including UBF ( D ), RRN3 ( E ) and RPA40 ( F ); IF assays were performed with HepG2 cells. Scale bars in the images represent 5 μm. G – I Representative images showing coimmunoprecipitation between EGR1 and components of the Pol Ⅰ transcription machinery in HeLa cells. Co-IP assays were performed using HeLa nuclear extract and an anti-EGR1 antibody, and proteins precipitated by the EGR1 antibody were detected by Western blot using antibodies against the indicated factors ( G ). The results of reciprocal co-IP assays are presented in ( H and I ). J – L Representative images showing the coimmunoprecipitation of EGR1 and components of the Pol Ⅰ transcription machinery in HepG2 cells. Co-IP assays were performed as described in ( G – I ). The input in ( G – L ) is equivalent to 2.5% of the samples used for each IP assay.

Article Snippet: Co-IP assays were performed using an anti-EGR1 antibody (SC-515830; Santa Cruz Biotech).

Techniques: Immunofluorescence, Co-Immunoprecipitation Assay, Western Blot