enzyme linked immunosorbent assay elisa kit  (R&D Systems)

Journal: Cell Reports Medicine

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

doi: 10.1016/j.xcrm.2026.102717

Figure Lengend Snippet: G9a inhibition potentiates the immunogenic effects of IFN-γ in HCC cells (A) Effect of CM272 and EZM8266 on IFN-γ-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 IFN-γ (75 U/mL) for another 24 h or with IFN-γ alone for 24 h. PM299L were treated with 400 nM, and HuH7 received 1 μM of CM272 ( n = 3). For EZM8266, cells were pretreated for 48 h with EZM8266 (5 mM) and then with IFN-γ (75 U/mL) for another 24 h, as indicated. CXCL10 protein levels were measured by ELISA in cells’ conditioned media ( n = 3). (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 IFN-γ and CM272 or EZM8266 as indicated in (A), and MHC-I levels were determined by FACS analysis ( n = 3). (C) ChIP analyses of H3K9me2 levels in the proximal promoter regions of CXCL10 and HLA-A genes in HuH7 cells treated with IFN-γ (75 U/mL) and CM272 (400 nM), as indicated in (A) ( n = 3). (D) Evaluation of the expression of transposable elements (TEs) and endogenous retroviral sequences (ERVs) by RNA-seq in NM53 murine HCC cells treated with IFN-γ, CM272, and their combination as indicated in (A). (E) Immunofluorescence analyses of dsRNA in PM299L HCC cells treated with CM272 (24 h) or EZM8266 (48 h). Representative images are shown. Scale bars, 10 μm. Right panel shows a control without primary antibody ( n = 3). Data are represented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. All the replicates represent biological replicates.

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 (DY492) and CXCL10 (DY466), both from R&D Systems, and an ELISA kit for human CXCL10 (550926) from BD Biosciences (Franklin Lanes, NJ, USA).

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

Journal: bioRxiv

Article Title: A multivalent peptide-polymer conjugate material mimics STING to therapeutically activate innate immune signaling

doi: 10.64898/2026.03.24.712780

Figure Lengend Snippet: ( A ) HEK293T reporter cells were treated with 6.0 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle, cells were imaged by confocal microscopy at 6 h post treatment to examine conjugate colocalization with target TBK1 (representative of N = 3 biological replicates). Scale bar is 10 μm. ( B ) HEK293T reporter cells were treated with 8.3 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle, Western blot was performed at 6 h post treatment to examine TBK1 and IRF3 phosphorylation (representative of N = 3 biological replicates). ( C ) IRF3 reporter signal relative to buffer treatment for HEK293T reporter cells pretreated for 6 h with TBK1 inhibitor MRT67307 (TBK1i) and then treated with 8.3 μg/mL STING or Scr conjugate delivered using TransIT-X2, measured 24 h post treatment (N = 3 biological replicates). ( D ) Western blot of STING and β-actin expression in ovarian cancer cell lines KURAMOCHI and A2780. ( E-F ) KURAMOCHI and A2780 ovarian cancer cell lines were treated with 5.0 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle or 50 μM STING agonist ADU-S100. ( E ) CXCL10 and ( F ) IFN-β in supernatant was measured by ELISA 24 h post treatment (N = 3 biological replicates). Replicates where analyte was below the limit of detection (LOD) are labeled as not detected (ND), no summary statistics were computed if any replicate was ND. ( G-I ) KURAMOCHI cells were treated with 5.0 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle or 50 μM ADU-S100, mRNA sequencing was performed at 6 h post treatment (N = 4 biological replicates). ( G ) Plot of Log2 fold change of STING conjugate or ADU-S100 treatment compared to Buffer, showing high correlation between treatments. Plot of Log2 fold change of Scr conjugate or ADU-S100 treatment compared to Buffer is displayed below as a control, showing greatly reduced correlation. The coefficient of determination R 2 for line of best fit is displayed. ( H ) Gene set enrichment analysis was performed on MSigDB Hallmark gene set, normalized enrichment and adjusted P value are displayed for the 10 gene sets significantly enriched ( P < .05) when comparing STING to Scr conjugate. Normalized enrichment and adjusted P values for the same 10 gene sets are displayed for the comparison of ADU-S100 to Buffer ( I ) Heatmap of gene expression for selected genes. Replicates where a given gene was not detected are labeled ND. Data represented as geometric mean ± SD.

Article Snippet: DuoSet ELISA kits (R&D Systems) for human CXCL10 (#DY266), human IFN-β (#DY814), mouse CXCL10 (#DY466), mouse IFN-β (#DY8234), mouse IL-6 (#DY406), mouse TNF-α (#DY410), and mouse IFN-γ (#DY485) were used with 1-Step TMB ELISA Substrate Solution (Thermo Scientific) following vendor instructions.

Techniques: Confocal Microscopy, Western Blot, Phospho-proteomics, Expressing, Enzyme-linked Immunosorbent Assay, Labeling, Sequencing, Control, Comparison, Gene Expression

Journal: bioRxiv

Article Title: A multivalent peptide-polymer conjugate material mimics STING to therapeutically activate innate immune signaling

doi: 10.64898/2026.03.24.712780

Figure Lengend Snippet: ( A-B ) Mice were dosed with 20 μg of STING conjugate delivered by LNP IP and serum was collected at 0, 1, 3, 6, 10, 24, and 50 h. Serum was analyzed to measure ( A ) STING conjugate concentration by Cy5 fluorescence (showing one phase exponential decay fit to data) and ( B ) CXCL10 concentration by ELISA (N = 3 mice). Conditions where analyte was below the limit of detection (LOD) are labeled as not detected (ND). ( C ) Mice were inoculated with 3×10 6 BPPNM cells IP and dosed with 20 μg of STING or Scr conjugate (N = 3 mice) delivered by LNP IP at 14 days after inoculation. ( D ) Omental tumor, ( E ) ascites, and ( F ) serum were collected 6 h after dosing. Concentrations of CXCL10, IFN-β, IL-6, TNF-α, and IFN-γ were measured by ELISA and are reported relative to total protein concentration in tumor and ascites ( D-E ) or relative to volume in serum ( F ). Conditions where analyte was below the LOD are labeled as ND. Average fold change increases in cytokine concentration for STING conjugate treatment compared to Scr conjugate are displayed. Where cytokine level was ND, a lower bound on the fold change was computed by setting all ND replicates as the LOD. Data represented as mean ± SD.

Article Snippet: DuoSet ELISA kits (R&D Systems) for human CXCL10 (#DY266), human IFN-β (#DY814), mouse CXCL10 (#DY466), mouse IFN-β (#DY8234), mouse IL-6 (#DY406), mouse TNF-α (#DY410), and mouse IFN-γ (#DY485) were used with 1-Step TMB ELISA Substrate Solution (Thermo Scientific) following vendor instructions.

Techniques: Concentration Assay, Fluorescence, Enzyme-linked Immunosorbent Assay, Labeling, Protein Concentration