rabbit polyclonal antibody against c jun ap 1  (Proteintech)

Journal: International immunopharmacology

Article Title: IL17A/F secreted by ASCT2-overexpression ovarian cancer cells contributes to immune escape through the suppression of natural killer (NK) cells cytotoxicity by the activation of c-JUN/ PTGS2 pathway.

doi: 10.1016/j.intimp.2025.114226

Figure Lengend Snippet: Fig. 5. Analysis of gene expression changes in NK cells co-cultured with ASCT2-overexpressing ovarian cancer cells. A: Volcano plot of differentially expressed genes, with significantly upregulated genes shown in red points, downregulated genes shown in blue points, and genes with no significant differential expression shown in gray points; the x-axis represents the fold change in gene expression between different groups, and the y-axis represents the statistical significance of the difference in gene expression. B: KEGG pathway enrichment plot of differentially expressed genes, with the x-axis representing the ratio of differentially expressed genes annotated to KEGG pathways to the total number of differentially expressed genes, and the y-axis representing KEGG pathways. C: Protein interaction plot of the top 100 differentially significant genes in NK cells. Genes shown in orange are upregulated, while genes shown in green are downregulated. D: Schematic diagram of the IL-17 signaling pathway. E, F, G: Detection of JUN and PTGS2 expression using RNA sequencing, qPCR, and Western blot analysis. OE vs NC, Comparison between NK cells co-cultured with ASCT2-overexpressing ovarian cancer cells and NK cells co-cultured with ASCT2-normal ovarian cancer cells. OE-NC, NK cells after co-culture with ovarian cancer cells with normal ASCT2 expression; OE-ASCT2, NK cells after co-culture with ASCT2-overexpressed ovarian cancer cells. The expression of JUN and PTGS2 in NK cells co-cultured with ASCT-2 overexpression was higher than that in NK cells co-cultured with normal ASCT-2 expression.

Article Snippet: The membrane was blocked in Tris-buffered saline (pH 7.4) containing 5 % skim milk and 0.1 % Tween-20, followed by overnight incubation at 4 ◦C with the primary antibody PTGS2 (66351-1-Ig, Proteintech, 1:4000)、 JUN (22114-1-AP, Proteintech Dilution: 1:1000) 、GADPH (GB15002, seivicebio Dilution: 1:2000) and subsequent incubation with the appropriate secondary antibody (AB 228338 Invitrogen, 1:3000) for 1 h. Protein bands were visualized using a Gel Imaging System (BIO-RAD, USA), and band intensities were quantified using ImageJ software.

Techniques: Gene Expression, Cell Culture, Quantitative Proteomics, Expressing, RNA Sequencing, Western Blot, Comparison, Co-Culture Assay, Over Expression

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Echinochrome A inhibits HMGB1-induced vascular smooth muscle cell migration by suppressing osteopontin expression

doi: 10.4196/kjpp.24.220

Figure Lengend Snippet: (A) Three lengths of the OPN promoter were individually constructed in a luciferase-based reporter vector to produce pGL3-OPN-2284 (full-length), pGL3-OPN-538 and pGL3-OPN-234 promoter constructs. The VSMCs were transiently transfected with these promoter constructs and an empty luciferase vector (pGL3) for 24 h, and then stimulated with HMGB1 (100 ng/ml) for 1 h. The differences in promoter activity between the control and HMGB1-stimulated cells were presented as the means ± SEMs of 6 independent experiments. **p < 0.01 vs . non-treated control. (B) Nucleotide sequence of the -538 ~ -234 region of the OPN promoter. The transcription factor binding sites were identified using TF Search software. The sequences of the potential binding sites for AP-1 and C/EBPβ in pLuc-OPN-538 were underlined. (C) The binding activity of AP-1 on the OPN promoter segment in HMGB1-treated VSMCs was assessed using a ChIP assay. IgG was used as the negative control. The relative binding intensities to the input signals were quantified and presented as the means ± SEMs of 6 independent experiments. **p < 0.01 vs . the value at 0 h. OPN, osteopontin; VSMCs, vascular smooth muscle cells; HMGB1, high-mobility group box 1; AP-1, activator protein 1; ChIP, chromatin immunoprecipitation.

Article Snippet: The activator protein 1 (AP-1) (10024-2-AP) antibody was purchased from Proteintech (Proteintech Group).

Techniques: Construct, Luciferase, Plasmid Preparation, Transfection, Activity Assay, Control, Sequencing, Binding Assay, Software, Negative Control, Chromatin Immunoprecipitation

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Echinochrome A inhibits HMGB1-induced vascular smooth muscle cell migration by suppressing osteopontin expression

doi: 10.4196/kjpp.24.220

Figure Lengend Snippet: (A) The effects of Ech A on the binding activity of AP-1 on the OPN promoter were assessed using a ChIP assay. IgG was used as the negative control. The relative binding intensity to input signals was quantified, and expressed as the means ± SEMs of 6 independent experiments. **p < 0.01 vs . non-treated control, # p < 0.05 and ## p < 0.01 vs . vehicle in the HMGB1-treated group. (B) Cells were transiently transfected with pLuc-OPN-538 constructs for 24 h, and then stimulated with HMGB1 (100 ng/ml) for 1 h in the presence of the indicated concentrations of Ech A (3 or 10 μM). The relative luciferase activities were presented as the means ± SEM of 6 independent experiments. **p < 0.01 vs . control, # p < 0.05 and ## p < 0.01 vs . vehicle in the HMGB1-treated group. (C) VSMCs were pretreated with Ech A (3 or 10 μM) for 24 h and then stimulated with HMGB1 (100 ng/ml) for 6 h and 48 h to determine mRNA and protein expression, respectively. GAPDH and β-actin were used as the internal controls for mRNA and protein expression, respectively. The relative band intensities were quantified, and expressed as the mean ± SEM of 6 independent experiments. **p < 0.01 vs . corresponding control, # p < 0.05 and ## p < 0.01 vs . corresponding value in vehicle of the HMGB1-treated group. OPN, osteopontin; VSMCs, vascular smooth muscle cells; HMGB1, high-mobility group box 1; AP-1, activator protein 1; ChIP, chromatin immunoprecipitation.

Article Snippet: The activator protein 1 (AP-1) (10024-2-AP) antibody was purchased from Proteintech (Proteintech Group).

Techniques: Binding Assay, Activity Assay, Negative Control, Control, Transfection, Construct, Luciferase, Expressing, Chromatin Immunoprecipitation