Journal: Technology in Cancer Research & Treatment

Article Title: Combining Radiation and anti-PD-L1 Enhances the Antitumor Activity in Colorectal Cancer via IFN-γ-Dependent Activation of STAT1

doi: 10.1177/15330338251406931

Figure Lengend Snippet: Immune activation in tumor and spleen. (A) Immunohistochemical staining for CD3 and CD8 in tumor sections (100 × and 400×) with integrated optical-density quantification (CD3 IOD + CD8 IOD); n = 4, mean ± SD; Representative fields were randomly selected from biological samples, with multiple technical replicates per sample. Statistical analysis: one-way ANOVA. (B) Flow cytometric analysis of splenic CD4 + and CD8 + T-cell subsets. First row of images: Heat map of CD45 + cells in each group of spleen cells. Values among the figure: Percentage of CD45 + cells in spleen cells. Second row image: Heat map of T cell subsets. The quadrants represent: Q2 CD3 + CD4 + , ie, CD4 + T cells; Q3 CD3 + CD4 + , ie, CD8 + T cells. The values among the image represent the percentage of the cell subset in CD45 + cells. (C) IFN-γ concentrations in tumor and spleen measured using ELISA (n = 4); Tumors were divided for different downstream assays: two frozen for cytokine analysis. ELISA results therefore reflect technical duplicates from available biological samples (n = 4, mean ± SD, technical replicates). Statistical analysis: one-way ANOVA.

Article Snippet: Therapeutic agents: anti-mouse PD-L1 antibody (clone 10F.9G2), recombinant mouse IFN-γ and Itacitinib (both MedChemExpress).

Techniques: Activation Assay, Immunohistochemical staining, Staining, Enzyme-linked Immunosorbent Assay

Journal: Technology in Cancer Research & Treatment

Article Title: Combining Radiation and anti-PD-L1 Enhances the Antitumor Activity in Colorectal Cancer via IFN-γ-Dependent Activation of STAT1

doi: 10.1177/15330338251406931

Figure Lengend Snippet: Function of the IFN-γ–JAK–STAT axis. (A) Immunohistochemistry for phospho-STAT1 in tumor tissues (100 × and 400×); n = 4, mean ± SD. Four representative fields were randomly selected from biological samples (technical replicates). Statistical analysis: one-way ANOVA. (B) Western blots showing expression of p-JAK1, p-STAT1, and cleaved caspase-3 in CT26.WT and MC38 cells under control, IR, IFN-γ, and IR + IFN-γ conditions. Grayscale densitometry for all Western blots has been performed using ImageJ, normalized to β-actin (n = 3 independent experiments). Bars represent mean ± SD. Statistical comparison was performed using an unpaired two-tailed t-test. (C) Apoptosis was quantified by Annexin V-FITC/PI staining in CT26.WT cells (n = 3 independent experiments per group). One-way ANOVA revealed a significant overall effect of treatment (P < 0.0001).

Article Snippet: Therapeutic agents: anti-mouse PD-L1 antibody (clone 10F.9G2), recombinant mouse IFN-γ and Itacitinib (both MedChemExpress).

Techniques: Immunohistochemistry, Western Blot, Expressing, Control, Comparison, Two Tailed Test, Staining

Journal: Technology in Cancer Research & Treatment

Article Title: Combining Radiation and anti-PD-L1 Enhances the Antitumor Activity in Colorectal Cancer via IFN-γ-Dependent Activation of STAT1

doi: 10.1177/15330338251406931

Figure Lengend Snippet: STAT1 inhibition assay. (A-B) Western blots and Annexin V/PI staining demonstrated that pharmacologic inhibition of STAT1 by Itacitinib abolished IFN-γ induced caspase-3 cleavage, and apoptosis in CT26.WT cells. Grayscale densitometry for all Western blots has been performed using ImageJ, normalized to β-actin (n = 3 independent experiments). Bars represent mean ± SD. Statistical comparison was performed using an unpaired two-tailed t-test.

Article Snippet: Therapeutic agents: anti-mouse PD-L1 antibody (clone 10F.9G2), recombinant mouse IFN-γ and Itacitinib (both MedChemExpress).

Techniques: Inhibition, Western Blot, Staining, Comparison, Two Tailed Test