Journal: iScience

Article Title: Loss of ZNF408 attenuates STING-mediated immune surveillance in breast carcinogenesis

doi: 10.1016/j.isci.2024.110276

Figure Lengend Snippet: ZNF408 activates STING1 expression and elicits the STING-IRF3 inflammatory response (A) DAPI and dsDNA staining of MCF-7 cells digested with 50 μg/mL DNase I. Scale bar, 20 μm. (B) MCF-7 cells were stably depleted ZNF408 with or without overexpressing STING1, or were stably overexpressed ZNF408 with or without depleting STING1. Cell lysates were collected and analyzed by western blotting with antibodies of ZNF408, STING1, IRF3, p-IRF3, P65, or p-P65 (left). The bands were quantified with ImageJ software. Each bar represents the mean ± SD for triplicate experiments. Mean data are normalized to β-actin (∗ p < 0.05) (right). (C) RT-qPCR analysis of representative cytokines activated by the cGAS/STING pathway in the engineered MCF-7 cells described previously. (D) ELISA analysis of IFNβ, CCL5, and CXCL10 in the medium collected from of the engineered MCF-7 cells described previously. In (B–D), each bar represents the mean ± SD for triplicate experiments (∗ p < 0.05, two-tailed t test). (E) Crystal violet staining of the indicated MCF-7 cells co-cultured with activated CD8 + T cells for 60 h (left). Histogram shows the relative intensity of cells (right). (F) Crystal violet staining images (left) and quantification of relative intensity (right) of MCF-7 cells 3 days after infection with HSV-1 at MOI 0.5. (G) RT-qPCR analysis of cellular HSV-1 genome loads in MCF-7 cells infected with HSV-1 (MOI = 0.5). Each bar represents the mean ± SD for triplicate experiments. Mean data are normalized to GAPDH (∗ p < 0.05). (H) MCF-7 cells depleted ZNF408 with or without overexpression of STING1 were split into 96-well plates and harvested at the indicated time points. Cell growth curves were measured with CCK-8 assays ( n = 6). The absorbance values were measured at 450 nm and normalized by subtracting the absorbance of the cultured medium alone. In (E–H), each bar represents the mean ± SD for triplicate experiments (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, two-way ANOVA and two-tailed t test). See also Figure S1 and Table S3 .

Article Snippet: CXCL10 human ELISA kit , Proteintech , Cat# KE00128.

Techniques: Expressing, Staining, Stable Transfection, Western Blot, Software, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Cell Culture, Infection, Over Expression, CCK-8 Assay

Journal: iScience

Article Title: Loss of ZNF408 attenuates STING-mediated immune surveillance in breast carcinogenesis

doi: 10.1016/j.isci.2024.110276

Figure Lengend Snippet:

Article Snippet: CXCL10 human ELISA kit , Proteintech , Cat# KE00128.

Techniques: Purification, Virus, Recombinant, Protease Inhibitor, Silver Staining, Bicinchoninic Acid Protein Assay, Transfection, Plasmid Preparation, cDNA Synthesis, Enzyme-linked Immunosorbent Assay, shRNA, Software