human il 8 precoated elisa kit  (Dakewe Biotech Co)

Journal: Immunity, Inflammation and Disease

Article Title: Ginkgolide B Alleviates Airway Inflammation in Hyperoxia Lung Injury

doi: 10.1002/iid3.70364

Figure Lengend Snippet: PAF stimulates neutrophils to be resident locally and secrete IL‐6. (A) The level of PAF in plasma from preterm neonates with BPD ( n = 8) was higher than that in non‐BPD group ( n = 8). (B) The level of PAF in BALF from preterm neonates with BPD ( n = 14) increased higher than that in non‐BPD group ( n = 9). (C) PAF level in BALF was positively correlated with oxygen supplement days in patients with BPD ( n = 14). (D–F) The percentages of CD66b + CD62L + in neutrophils from the blood of healthy neonates at birth decreased sharply after PAF (100 ng/mL) stimulation at different time points. (G) The level of soluble CD62L in the plasma of patients with BPD ( n = 11) or without BPD ( n = 15). (H–K) Relative mRNA expressions of IL6 , LCN2 , IL10 , and MMP12 were examined in neutrophils after PAF (100 ng/mL) stimulation for 4 h. (L) PAF induced neutrophils to form NETs (MPO represents neutrophil activation, and the red arrows indicate the presence of free DNA outside the cells, which signifies the formation of NETs). (M) Clustering of inflammatory cytokine levels in BALF from preterm neonates with and without BPD. (N) Differential inflammatory cytokines in BALF from patients with and without BPD were displayed in heatmap. (O–R) The levels of IL‐6, IL‐8, and GM‐CSF were tested higher in BALF of BPD patients ( n = 9–26) postnatal 1 week than those of non‐BPD ( n = 6–18), except for GRO‐α by ELISA. Each experiment was independently repeated at least three times.

Article Snippet: Human IL‐8 Precoated ELISA Kit , DAKEWE , 1110802 , .

Techniques: Clinical Proteomics, Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Biological Chemistry

Article Title: Epstein-Barr virus suppresses N 6 -methyladenosine modification of TLR9 to promote immune evasion

doi: 10.1016/j.jbc.2024.107226

Figure Lengend Snippet: TLR9 downstream signaling pathways and cytokine expression are regulated by METTL3. A , cellular m 6 A modification levels were assayed in BJAB-shNC and BJAB-shMETTL3 cells by Dot blot analysis. B , the expression levels of TLR9 and METTL3 were determined by Western blotting in BJAB-shNC and BJAB-shMETTL3 cells. C , BJAB-shNC and BJAB-shMETTL3 cells were treated with ActD (5 μg/ml) for 0, 3, and 6 h; TLR9 mRNA levels were measured by RT-qPCR. D , the indicated TLR9 signaling proteins were detected by cytoplasmic and nuclear proteins isolation. GAPDH and Histone H3 were used for cytoplasm and nucleus protein loading controls, respectively. E , BJAB cells were treated with STM2457(10 μM) and CpG-ODN2006 (5 μM) upon 50 MOI EBV infection, respectively. EBV genomes copy number was determined by qPCR. F , primary B cells were treated with STM2457(10 μM) and CpG-ODN2006(5 μM) with or without 50MOI EBV infection for appointed time. The secretion of IFN-α and CXCL10 were assayed by ELISA. These data are shown as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ns, not significant. EBNA1, Epstein-Barr nuclear antigen 1; EBV, Epstein-Barr virus; IFN, interferon; m 6 A, N 6 -methyladenosine; MOI, multiplicity of infection; ODN, oligodeoxynucleotide; RT-qPCR, reverse transcription-quantitative PCR; TLR9, toll-like receptor 9.

Article Snippet: ELISA kits were obtained from Dakewe: Human IFN-α Precoated ELISA Kit (Cat#:1110012), Human IP-10 Precoated ELISA Kit (Cat#:1117452), Human IgA Precoated ELISA Kit (Cat#:1128172), Human IgM Precoated ELISA Kit (Cat#:1128182), Human IgG Precoated ELISA Kit(Cat#:1128162), Human IL-6 Precoated ELISA Kit(Cat#:1110602), Human IL-8 Precoated ELISA Kit (Cat#:1110802).

Techniques: Protein-Protein interactions, Expressing, Modification, Dot Blot, Western Blot, Quantitative RT-PCR, Isolation, Infection, Enzyme-linked Immunosorbent Assay, Virus, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: Epstein-Barr virus suppresses N 6 -methyladenosine modification of TLR9 to promote immune evasion

doi: 10.1016/j.jbc.2024.107226

Figure Lengend Snippet: STM2457 inhibits CpG-ODN-induced B cell proliferation, Ig secretion and cytokine expressions. A , BJAB cells were treated with STM2457 for 96 h. Cellular m 6 A modification levels were assayed by dot blot analysis. TLR9 signaling molecules were assayed by Western blotting. B , gene set difference between the TLR9 high and low human immune cells was revealed by GSEA. The top gene set enriched in TLR9 high group comparing to low group is listed. C–E , PBMCs from healthy donors were stained with CFSE dyes and cultured with anti-IgM (10 mM), CD40L (1 mM), and IL-4 (10 nM), costimulated with STM2457(10 μM) or ODN-2006(5 μM) for 0 and 6 days. DMSO was used as a control. Only CD19+ B cells stained CFSE dyes were gated by flow cytometry analysis ( C and D ). E , histogram shows the percentage of B cells dividing under different conditions. F and G , fresh PBMCs cultured with anti-IgM, CD40L, IL-4, and costimulated with STM2457(10 μM) or ODN-2006(5 μM) for 8 days were collected to detect Ig-producing cells ( F ) and cytokine level in supernatants by ELISA analysis ( G ). H and I , BJAB-shRNA cells were transfected with HA-TLR9 plasmids for 48 h. Then, the levels of Ig ( H ) and cytokine ( I ) in supernatants were detected by ELISA analysis. J , model diagram demonstrates the role of STM2457 in the regulation of B-cell proliferation and cytokines secretion. Experiments were independently repeated three times, and results are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ns, not significant, compared with the control group. CFSE, carboxyfluorescein succinimidyl ester; DMSO, dimethylsulfoxide; GSEA, gene set enrichment analysis; Ig, immunoglobulin; IL, interleukin; m 6 A, N 6 -methyladenosine; ODN, oligodeoxynucleotide; PBMC, peripheral blood mononuclear cell; TLR9, toll-like receptor 9.

Article Snippet: ELISA kits were obtained from Dakewe: Human IFN-α Precoated ELISA Kit (Cat#:1110012), Human IP-10 Precoated ELISA Kit (Cat#:1117452), Human IgA Precoated ELISA Kit (Cat#:1128172), Human IgM Precoated ELISA Kit (Cat#:1128182), Human IgG Precoated ELISA Kit(Cat#:1128162), Human IL-6 Precoated ELISA Kit(Cat#:1110602), Human IL-8 Precoated ELISA Kit (Cat#:1110802).

Techniques: Modification, Dot Blot, Western Blot, Staining, Cell Culture, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, shRNA, Transfection