duoset elisa kit  (R&D Systems)

Journal: bioRxiv

Article Title: Selective JAK Inhibition Reveals Paradoxical and Hierarchical Control of interferon-γ-driven Autoimmunity in AIRE Deficiency

doi: 10.64898/2026.03.05.709894

Figure Lengend Snippet: Untreated Air e −/− mice and Air e −/− mice treated for four weeks with selective JAK1i, JAK2i, or JAK3i were analyzed. Lungs were processed for intracellular cytokine staining, qPCR, ELISA, and immunoblot analyses. ( A-B ) Representative flow cytometry plots showing IFN-γ production by CD4 + and CD8 + T cells. ( C-D ) Frequency (of total CD4 + and CD8 + T cells) and absolute numbers of IFN-γ + CD4 + and IFN-γ + CD8 + T cells in the lung. ( E-F ) Relative Ifng mRNA expression and IFN-γ protein concentrations in lung homogenates. ( G ) Relative Stat1 mRNA expression. ( H ) Representative immunoblots of phospho-STAT1 (pSTAT1), total STAT1, and GAPDH. ( I-J ) Quantification of total STAT1 and phospho-STAT1 normalized to GAPDH. For IFN- γ + CD4 + and IFN-γ + CD8 + T cells: n = 9-14 mice per group from four independent experiments. For Ifng and Cxcl9 mRNA: n = 15-22 mice per group from four independent experiments. For Stat1 _mRNA: n = 10-17 mice per group from three independent experiments. For CXCL9 protein: n = 5-10 mice per group from two independent experiments. For STAT1 and pSTAT1 immunoblots: n = 15-22 mice per group from four independent experiments). Statistical analyses were performed using one-way ANOVA or Kruskal-Wallis test with Dunn’s multiple comparisons to the untreated Air e −/− mice. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: A DuoSet® ELISA kit (catalog no. DY492-05; R&D Systems, USA) was used to measure CXCL9 and a high-sensitivity ELISA kit was used for IFN-γ (catalog no. 88-8314-88; Invitrogen, USA).

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Flow Cytometry, Expressing

Journal: bioRxiv

Article Title: Histone methyl-transferase G9a inhibition boosts the efficacy of immune checkpoint inhibitors in experimental hepatocellular carcinoma

doi: 10.1101/2025.07.08.663670

Figure Lengend Snippet: (A) Effect of CM272 and EZM8266 on IFNγ-triggered CXCL10 production in murine (PM299L) and human (HuH7) HCC cells. Cells were treated with CM272 for 48 h or with CM272 for 24 h and then with INFγ (75 U/mL) for another 24 h, or with INFγ alone for 24 h. PM299L were treated with 400 nM and HuH7 received 1 µM of CM272. For EZM8266, cells were pretreated for 48 h with EZM8266 (5 µM) and then with IFNγ (75 U/mL) for another 24 h as indicated. CXCL10 protein levels were measured by ELISA in cells’ conditioned media. (B) Effect of G9a inhibition with CM272 or EZM8266 on the expression of MHC class I complex protein (MHC-I) on the surface of PM299L cells. Cells were treated with IFNγ and CM272 or EZM8266 as indicated in panel A, and MHC-I levels were determined by FACS analysis. (C) ChIP analyses of H3K9me2 levels in the proximal promoter regions of CXCL10 and HLA-A genes in HuH7 cells treated with IFNγ (75 U/mL) and CM272 (400 nM) as indicated in panel A. (D) Evaluation of the expression of Transposable elements (TEs) and endogenous retroviral sequences (ERVs) by RNA-seq in NM53 murine HCC cells treated with IFNγ, CM272 and their combination as indicated in panel A. (E) Immunofluorescence analyses of dsRNA in PM299L HCC cells treated with CM272 (24 h) or EZM8266 for (48 h) h. Right panel shows a control without primary antibody. * p<0.05, ** p<0.01, *** p<0.001.

Article Snippet: CXCL9 and CXCL10 concentrations were measured in culture supernatants collected at the end of the incubation periods using commercial ELISA kits for mouse (CXCL9: R&D Systems, DY492; CXCL10: R&D Systems, DY466) and human cell lines (CXCL10: BD Biosciences, 550926).

Techniques: Enzyme-linked Immunosorbent Assay, Inhibition, Expressing, Retroviral, RNA Sequencing, Immunofluorescence, Control

Journal: BMC biotechnology

Article Title: Parabacteroides distasonis promotes CXCL9 secretion of tumor-associated macrophages and enhances CD8 + T cell activity to trigger anti-tumor immunity against anti-PD-1 treatment in non-small cell lung cancer mice.

doi: 10.1186/s12896-025-00963-9

Figure Lengend Snippet: Fig. 6 P. distasonis enhanced TAM-mediated CXCL9 secretion. (A) Flow cytometry detection of the proportion of CXCL9+F4/80+ macrophages in mouse tumor tissues. *P < 0.05 vs. PBS. (B) Western blot assessment of CXCL9 protein levels in BMDM. (C) ELISA analysis of the secretion level of CXCL9 in BMDM supernatant. *P < 0.05 vs. Control

Article Snippet: TNF-α (CSB-E04741m, CUSABIO), IL-6 (CSB-E04639m, CUSABIO), IL-1β (CSB-E08054m, CUSABIO), CXCL9 (CSB-EL006252MO, CUSABIO), IFN-γ (CSB-E04578m, CUSABIO), Perforin (CSB-E13429m, CUSABIO), and granzyme B (GZMB, CSB-E08720m, CUSABIO) kits were conducted to determine TNF-α, IL-6, IL-1β and CXCL9 levels in BMDM supernatant and secretion of IFN-γ, TNF-α, Perforin and GZMB in primary CD8+T cell supernatant of spleen.

Techniques: Flow Cytometry, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: BMC biotechnology

Article Title: Parabacteroides distasonis promotes CXCL9 secretion of tumor-associated macrophages and enhances CD8 + T cell activity to trigger anti-tumor immunity against anti-PD-1 treatment in non-small cell lung cancer mice.

doi: 10.1186/s12896-025-00963-9

Figure Lengend Snippet: Fig. 7 P. distasonis enhanced CD8+T cell activity by secreting CXCL9 from macrophages in vivo. (A) Flow cytometry determination of positive proportion of CXCR3 in CD8+T cells in mouse tumor tissue. *P < 0.05 vs. PBS. (B) Tumor images. (C) Tumor volume growth curve. (D) Tumor weight. (E) IHC staining of Ki67 expression in mouse tumor tissue. Scale bar = 100 μm (100×), Scale bar = 25 μm (400×). (F and G) Flow cytometry determination of positive propor tion of GZMB and IFN-γ in CD8+T cells in mouse tumor tissue. *P < 0.05 vs. αPD-1, &P < 0.05 vs. αPD-1 + P. distasonis

Article Snippet: TNF-α (CSB-E04741m, CUSABIO), IL-6 (CSB-E04639m, CUSABIO), IL-1β (CSB-E08054m, CUSABIO), CXCL9 (CSB-EL006252MO, CUSABIO), IFN-γ (CSB-E04578m, CUSABIO), Perforin (CSB-E13429m, CUSABIO), and granzyme B (GZMB, CSB-E08720m, CUSABIO) kits were conducted to determine TNF-α, IL-6, IL-1β and CXCL9 levels in BMDM supernatant and secretion of IFN-γ, TNF-α, Perforin and GZMB in primary CD8+T cell supernatant of spleen.

Techniques: Activity Assay, In Vivo, Flow Cytometry, Immunohistochemistry, Expressing

Journal: BMC biotechnology

Article Title: Parabacteroides distasonis promotes CXCL9 secretion of tumor-associated macrophages and enhances CD8 + T cell activity to trigger anti-tumor immunity against anti-PD-1 treatment in non-small cell lung cancer mice.

doi: 10.1186/s12896-025-00963-9

Figure Lengend Snippet: Fig. 8 P. distasonis promoted the secretion of CXCL9 by TAM and enhanced activity of CD8+T cells, triggering anti-tumor immunity against anti-PD-1 therapy in NSCLC mice

Article Snippet: TNF-α (CSB-E04741m, CUSABIO), IL-6 (CSB-E04639m, CUSABIO), IL-1β (CSB-E08054m, CUSABIO), CXCL9 (CSB-EL006252MO, CUSABIO), IFN-γ (CSB-E04578m, CUSABIO), Perforin (CSB-E13429m, CUSABIO), and granzyme B (GZMB, CSB-E08720m, CUSABIO) kits were conducted to determine TNF-α, IL-6, IL-1β and CXCL9 levels in BMDM supernatant and secretion of IFN-γ, TNF-α, Perforin and GZMB in primary CD8+T cell supernatant of spleen.

Techniques: Activity Assay