Journal: iScience

Article Title: DPPA inhibits melanoma by targeting angiogenesis through activating autocrine IFN-γ-CXCL9/10/11-CXCR3 axis in vascular endothelial cells

doi: 10.1016/j.isci.2026.116309

Figure Lengend Snippet: DPPA suppresses angiogenesis through the activation of the IFN-γ-CXCL9/10/11-CXCR3 axis in vascular endothelial cells (A) Angiogenesis PCR array for human dermal microvascular endothelial cells (HDMECs) treated with DMSO and DPPA at a concentration of 10 μM for 48 h. The relative expression levels were calculated as the log2 fold change, and the differentially expressed genes were selected on the basis of a log2 ≤ −2 or ≥2. (B) KEGG classification of differentially expressed genes in A. HDMECs were treated with DMSO and DPPA (10 μM) for 48 h, and RNAs and proteins were collected to analyze the expression of the IFN-γ-CXCL9/10/11 axis at both the transcriptional and protein levels using RT-qPCR (C) and western blotting (D) assays. n = 3 technical replicates from 3 biological replicates for each group. (E) 3D models of docking poses for DPPA and receptors (including IFN-γ, CXCL9, CXCL10, and CXCL11) predicted by Autodock Vina Tools. (F) 2D model of the interaction between DPPA and IFN-γ. HDMECs were treated with IFN-γ for 48 h, after which the RNAs and proteins were collected and subjected to RT-qPCR (G) and western blotting (H) to quantify the expression levels of CXCL9, CXCL10, CXCL11, p65, and pp65. β-actin was served as an internal control. (G) n = 3 technical replicates from 3 biological replicates for each group. (H) n = 3 biological replicates for each group. (I) HDMECs were treated with NF-κB inhibitor (NF-κB-IN-11) for 48 h, and RNAs were collected to analyze the expression of the IFN-γ at the transcriptional levels using qRT-PCR. β-actin was served as an internal control. n = 3 biological replicates for each group. Cell migration (J) and tube formation on Matrigel (K) of HDMECs treated with DMSO, DPPA, or DPPA with CXCL9/10/11-CXCR3 axis-blocking neutralizing antibodies. n = 3 technical replicates from 3 biological replicates for each group. Scale bars: 100 μm. Data are presented as mean ± SD. Significant effects: p values were calculated using Student’s t test (two-tailed unpaired t test) for C, D, G, H, and I, and one-way ANOVA for J and K. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 and ns: p > 0.05 (compared with the control/DMSO group).

Article Snippet: Human dermal microvascular endothelial cells (HDMECs, Procell system, Wuhan, China) were maintained in endothelial cell growth medium supplemented with growth supplements (EGM, CC-3124, Lonza), and have been authenticated by Procell system using CD31 immunofluorescence (IF) staining and been tested for mycoplasma contamination.

Techniques: Activation Assay, Concentration Assay, Expressing, Quantitative RT-PCR, Western Blot, Control, Migration, Blocking Assay, Two Tailed Test