mouse ifn-beta quantikine elisa kit  (Bio-Techne corporation)

Journal: Nature Communications

Article Title: Triple targeting of STING, TGF-β, and PD-L1 boosts CXCL16–CXCR6 signaling for potent antitumor response

doi: 10.1038/s41467-026-69456-3

Figure Lengend Snippet: A Tumor growth curves of EMT-6-bearing mice treated with control IgG (CTL) or YM101. Tumors were collected on day 19 ( n = 8 for CTL; n = 9 for YM101). B Tumor growth inhibition (TGI) of individual EMT-6 tumors; TGI < 50% was defined as nonresponder, and TGI ≥ 50% as responder. C GSEA of the GO term “innate immune response” comparing YM101 responders versus nonresponders in EMT-6 tumors. D Top enriched immune/cytokine GO terms ranked by normalized enrichment score (NES); all terms shown have FDR < 0.05. E Tumor growth curves of CT26-bearing mice treated with CTL or YM101. Tumors were collected on day 18 ( n = 6 for CTL; n = 15 for YM101). F TGI of individual CT26 tumors. G , H GSEA of the GO terms “innate immune response” and “response to IFN-β” comparing YM101 responders versus nonresponders in CT26 tumors. I Overlap of upregulated DEGs (UpDEGs) in YM101-responsive EMT-6 and CT26 tumors; representative genes are listed. J Functional enrichment analysis of overlapping upregulated DEGs. K Intratumoral cytokine profile in EMT-6 tumors from an independent cohort after YM101 treatment ( n = 6 for responders; n = 5 for nonresponders). L , O Representative tumor images from EMT-6 and CT26 models treated as indicated. M , P Tumor growth curves of EMT-6 and CT26 models ( n = 8 mice per group). N , Q Kaplan–Meier survival curves for EMT-6 and CT26 models ( n = 8 mice per group). A , E Data are mean ± SD. The P values shown were calculated using two-tailed unpaired t-tests. M , P Data are mean ± SD. Statistical comparisons were performed using one-way ANOVA followed by Holm–Šídák’s multiple comparisons test. N , Q P values were determined by the log-rank test. Statistical significance: * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Source data and exact P values are provided in the file.

Article Snippet: Total protein concentrations in tumor tissue homogenates were measured using a BCA protein assay kit, and cytokine levels were quantified using an IFN-β ELISA kit (MIFNB0, R&D Systems) and multiplex fluorescence-encoded bead-based assays (741043 and 740451, BioLegend).

Techniques: Control, Inhibition, Functional Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Triple targeting of STING, TGF-β, and PD-L1 boosts CXCL16–CXCR6 signaling for potent antitumor response

doi: 10.1038/s41467-026-69456-3

Figure Lengend Snippet: A Heatmap showing CXCL16 mRNA and protein levels in human macrophages (Mø) and dendritic cells (DC), and CXCL16-Luc reporter activity in HEK293 cells, under the following conditions: control (CTL), MSA-2, TGF-β1, MSA-2 + TGF-β1 (M + T), MSA-2 + TGF-β1 + α-TGF-β (M + T + α-T), and MSA-2 + TGF-β1 + α-PD-L1 (M + T + α-P). Data are presented as fold change (FC) relative to CTL ( n = 3 biological replicates). B STAT1 ChIP-seq tracks from three human macrophage datasets (GEO accessions GSM1057011 - GSM1057013 ), visualized via cistrome data browser and aligned to the CXCL16 locus (GRCh38/hg38). C Correlation analysis of CXCL16 expression with STAT1 and TMEM173 expression in the TCGA database. Correlations were calculated using Spearman’s rank correlation in TIMER2.0 tool. Each point represents a cancer type. D Schematic diagram of the CXCL16 promoter showing potential STAT1 binding sites. E Relative luciferase activity of CXCL16 promoter reporters after 24 h of MSA-2 treatment ( n = 3 biological replicates). F ChIP-qPCR analysis demonstrating STAT1 enrichment at the CXCL16 promoter in human Mø treated with MSA-2 for 48 h. Bars represent mean ± SD ( n = 3 biological replicates). G , H Heatmap of CXCL16 and IFN-β protein levels in human Mø and DC after treatment ( n = 3 biological replicates). I Western blot analysis of STING signaling activation, JAK1-STAT1 activation, and CXCL16 expression in human Mø and DC. J Heatmap showing IFN-β and CXCL16 protein expression in human Mø and DC ( n = 3 biological replicates). K Violin plot of Cxcl16 expression across various cell types in the tumor microenvironment. L Schematic of the experimental design to assess the dependence of M + Y synergy on intact JAK1-STAT1 signaling in myeloid cells. M Representative images of MC38 tumors. N Tumor growth curves showing mean tumor volume ± SD for MC38 tumors ( n = 6 per group). O Kaplan–Meier survival curve for MC38 tumor-bearing mice ( n = 6 per group). F Was analyzed using the two-tailed unpaired t -test. N Was analyzed using one-way ANOVA followed by Holm–Šídák’s multiple comparisons test. O Was analyzed by the log-rank test. In vitro results are representative of three independent experiments. Source data and exact P values are provided in the file.

Article Snippet: Total protein concentrations in tumor tissue homogenates were measured using a BCA protein assay kit, and cytokine levels were quantified using an IFN-β ELISA kit (MIFNB0, R&D Systems) and multiplex fluorescence-encoded bead-based assays (741043 and 740451, BioLegend).

Techniques: Activity Assay, Control, ChIP-sequencing, Expressing, Binding Assay, Luciferase, ChIP-qPCR, Western Blot, Activation Assay, Two Tailed Test, In Vitro

Journal: Advanced Science

Article Title: Immunometabolic Rewiring of Dendritic Cells to Overcome Glutamine‐Driven Immune Suppression in Colorectal Cancer

doi: 10.1002/advs.202513986

Figure Lengend Snippet: T26 enhances antitumor immunity through DC activation. (A) Experimental workflow for analyzing immune cell populations. C57BL/6 and BALB/c mice were subcutaneously inoculated with 1 × 10 6 MC38 and CT26 cells, respectively, into the right flank on day 0. Once tumors reached approximately 80 mm 3 , mice were randomly assigned to receive either JHU083 or MSA‐2 at 1 mg kg −1 day −1 or T26 at 1.5 mg kg −1 day −1 from day 7 to 11. After treatment, tumors, spleens, and draining lymph nodes were harvested for flow cytometry analysis. (B) Experimental design for ELISA quantification of pro‐inflammatory cytokines in serum and tumors in MC38 or CT26 bearing mice following PBS, JHU083, MSA‐2, or T26 treatment. (C) Flow cytometry analysis of mature DCs (CD80⁺CD86⁺) in MC38 tumors ( n = 3). (D) Flow cytometry analysis of CD8⁺ T cells (CD3⁺CD8⁺) in MC38 tumors ( n = 3). (E) Flow cytometry analysis of GZMB expression in tumor‐infiltrating CD8⁺ T cells in MC38 tumors ( n = 3). (F) Flow cytometry analysis of mature DCs (CD80⁺CD86⁺) in spleens ( n = 3). (G) Flow cytometry analysis of CD8⁺ T cells (CD3⁺CD8⁺) in spleens ( n = 3). (H) Flow cytometry analysis of mature DCs (CD80⁺CD86⁺) in TDLNs ( n = 3). (I–M) Tumor and serum levels of IFN‐α, IFN‐β, IFN‐γ, TNF‐α, IL‐6 in MC38‐bearing mice ( n = 3). (N) C57BL/6 mice with MC38 tumor were pretreated (i.v.) with either 5 mg per mouse of Cytc or PBS before other treatment and maintained the injection every other day for two weeks to deplete DCs. (O,P) Tumor growth of MC38 tumors in C57BL/6 mice with different treatment ( n = 6). Error bars represent means ± SD. Differences between groups were tested using one‐way ANOVA followed by Tukey's multiple comparisons test, or unpaired Student's t ‐test.

Article Snippet: The following ELISA kits were used: IFN‐α (ThermoFisher, BMS6027), IFN‐β (R&D Systems, MIFNB0), IL‐6 (Invitrogen, 88‐7064‐88), TNF‐α (Invitrogen, 88‐7324‐88), IFN‐γ (Invitrogen, 88‐7314‐88), HMGB1 (ThermoFisher, EEL102).

Techniques: Activation Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Injection