Journal: Frontiers in Oncology

Article Title: COL8A1 Promotes NSCLC Progression Through IFIT1/IFIT3-Mediated EGFR Activation

doi: 10.3389/fonc.2022.707525

Figure Lengend Snippet: COL8A1 stimulates IFIT1 and IFIT3 expression and EGFR activity in NSCLC cells. (A) RNA-seq data showing dysregulated genes in COL8A1-silenced NSCLC cells compared with shCtrl cells. (B) Gene set enrichment analysis (GSEA) of genes regulated in COL8A1-silenced and control cells. (C, D) qRT-PCR analysis of dysregulated genes in COL8A1-silenced and COL8A1-overexpressing cells. (E, F) Western blot evaluation of the effects of COL8A1-silencing and COL8A1-overexpression on the IFIT pathway and EGFR activity in NSCLC cells. (G, H) Correlation analysis between COL8A1 and IFIT1 or IFIT3 expression. *p < 0.05. **p < 0.01. ***p < 0.001.

Article Snippet: Primary antibodies against p-EGFR (#3777S, CST), EGFR (#2085, CST), p-AKT (#4060, CST), AKT (#4821, CST), IFIT-1 (23247-1-AP, Proteintech), IFIT-3 (15201-1-AP, Proteintech), and β-actin (#4970, CST) were applied in this study.

Techniques: Expressing, Activity Assay, RNA Sequencing, Control, Quantitative RT-PCR, Western Blot, Over Expression

Journal: Frontiers in Oncology

Article Title: COL8A1 Promotes NSCLC Progression Through IFIT1/IFIT3-Mediated EGFR Activation

doi: 10.3389/fonc.2022.707525

Figure Lengend Snippet: IFIT1 and IFIT3 knockdown effectively suppresses proliferation, migration, and apoptosis of NSCLC cells. (A, B) Effects of IFIT1 and IFIT3 knockdown on EGFR and AKT activity determined by western blotting. (C, D) Cell viability assays performed in IFIT1-knockdown H1299 and H1975 cells. (E, F) Cell viability assays performed in IFIT3-knockdown H1299 and H1975 cells. (G, H) Colony formation assessed in siCtrl-, siIFIT1-1, and siIFIT3-1-treated H1299 (G) and H1975 (H) cells. (I) Effects of IFIT1 and IFIT3 knockdown on apoptosis assessed by flow cytometry. (J) Effects of IFIT1 and IFIT3 knockdown on cell invasion and migration assessed by transwell assay. (K, L) H1299 and H1975 cells transfected with COL8A1, siIFIT1, and siIFIT3 were subjected to cell viability (K) and colony formation analysis (L) . **p < 0.01. ***p < 0.001.

Article Snippet: Primary antibodies against p-EGFR (#3777S, CST), EGFR (#2085, CST), p-AKT (#4060, CST), AKT (#4821, CST), IFIT-1 (23247-1-AP, Proteintech), IFIT-3 (15201-1-AP, Proteintech), and β-actin (#4970, CST) were applied in this study.

Techniques: Knockdown, Migration, Activity Assay, Western Blot, Flow Cytometry, Transwell Assay, Transfection

Journal: Frontiers in Oncology

Article Title: COL8A1 Promotes NSCLC Progression Through IFIT1/IFIT3-Mediated EGFR Activation

doi: 10.3389/fonc.2022.707525

Figure Lengend Snippet: COL8A1 contributes to NSCLC progression by activating EGFR in vivo . BALB/c nude mice were subcutaneously transplanted with COL8A1-silenced NSCLC cells. Mice were sacrificed 5 weeks after implantation. (A) Macroscopic images of xenografted tumors. (B) Tumor growth curves. (C) Tumor weight was determined immediately after sacrificing the mice. (D) Tumor tissues were subjected to western blotting analysis with the indicated antibodies. ***p < 0.001.

Article Snippet: Primary antibodies against p-EGFR (#3777S, CST), EGFR (#2085, CST), p-AKT (#4060, CST), AKT (#4821, CST), IFIT-1 (23247-1-AP, Proteintech), IFIT-3 (15201-1-AP, Proteintech), and β-actin (#4970, CST) were applied in this study.

Techniques: In Vivo, Western Blot

Journal: Journal of Inflammation Research

Article Title: Downregulation of IFIT3 Relieves the Inflammatory Response in Ulcerative Colitis via Selectively Regulating Macrophage M1 Polarization and the STAT1/2 Signaling Pathway

doi: 10.2147/JIR.S542033

Figure Lengend Snippet: IFIT3 was upregulated in colon tissues of DSS-induced UC mice. ( A ) Representative images of H&E-stained colon tissues from control and DSS-induced UC mice. Scale bar = 100 μm. ( B ) Protein levels of IFIT3 in colon tissues from the two mouse groups. ( C ) mRNA levels of IFIT3, TNF-α, IL-1β , and IL-6 in colon tissues from the two mouse groups. ( D ) Representative images of double immunofluorescence staining for IFIT3 and CD68 in colon tissues from the two mouse groups, and quantification of co-expressing IFIT3 and CD68 macrophages at 400x magnification. Scale bar = 100 μm. Data was presented as mean ± SD (n = 5). * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Following antigen retrieval and blocking, the tissue sections were incubated with primary antibodies against CD68 (1:500; #28058-1-AP, Proteintect) and IFIT3 (1:500; #15201-1-AP, Proteintech) at 4 °C overnight.

Techniques: Staining, Control, Double Immunofluorescence Staining, Expressing

Journal: Journal of Inflammation Research

Article Title: Downregulation of IFIT3 Relieves the Inflammatory Response in Ulcerative Colitis via Selectively Regulating Macrophage M1 Polarization and the STAT1/2 Signaling Pathway

doi: 10.2147/JIR.S542033

Figure Lengend Snippet: IFIT3 knockdown repressed macrophage M1 polarization by inhibiting the STAT1 pathway in vitro. ( A and B ) mRNA and protein levels of IFIT3 in untreated THP‐1 macrophages or treated with 100 ng/mL LPS or 100 ng/mL LPS + 20 ng/mL IFN‐γ or 20 ng/mL IL‐4. ( C ) mRNA levels of IFIT3 in 100 ng/mL LPS-stimulated macrophages treated with siNC or two siRNAs targeting IFIT3 (siIFIT3-1 and siIFIT3-2). ( D ) mRNA levels of TNF-α, IL-1β and IL-6 in 100 ng/mL LPS-stimulated macrophages treated with siNC and siIFIT3-2. ( E ) Protein levels of STAT1, pSTAT1, STAT2, pSTAT2 and IFIT3 in each group. ( F ) Representative gating and the co-expression of CD86 and CD68 in untreated (M0) and 100 ng/mL LPS-stimulated macrophages (M1) by flow cytometry. ( G ) Representative gating and the co-expression of CD68 and CD86 in 100 ng/mL LPS-stimulated macrophages treated with siNC and siIFIT3-2 by flow cytometry. Data was presented as mean ± SD from 3 independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Following antigen retrieval and blocking, the tissue sections were incubated with primary antibodies against CD68 (1:500; #28058-1-AP, Proteintect) and IFIT3 (1:500; #15201-1-AP, Proteintech) at 4 °C overnight.

Techniques: Knockdown, In Vitro, Expressing, Flow Cytometry

Journal: Frontiers in Oncology

Article Title: COL8A1 Promotes NSCLC Progression Through IFIT1/IFIT3-Mediated EGFR Activation

doi: 10.3389/fonc.2022.707525

Figure Lengend Snippet: COL8A1 stimulates IFIT1 and IFIT3 expression and EGFR activity in NSCLC cells. (A) RNA-seq data showing dysregulated genes in COL8A1-silenced NSCLC cells compared with shCtrl cells. (B) Gene set enrichment analysis (GSEA) of genes regulated in COL8A1-silenced and control cells. (C, D) qRT-PCR analysis of dysregulated genes in COL8A1-silenced and COL8A1-overexpressing cells. (E, F) Western blot evaluation of the effects of COL8A1-silencing and COL8A1-overexpression on the IFIT pathway and EGFR activity in NSCLC cells. (G, H) Correlation analysis between COL8A1 and IFIT1 or IFIT3 expression. *p < 0.05. **p < 0.01. ***p < 0.001.

Article Snippet: Primary antibodies against p-EGFR (#3777S, CST), EGFR (#2085, CST), p-AKT (#4060, CST), AKT (#4821, CST), IFIT-1 (23247-1-AP, Proteintech), IFIT-3 (15201-1-AP, Proteintech), and β-actin (#4970, CST) were applied in this study.

Techniques: Expressing, Activity Assay, RNA Sequencing, Control, Quantitative RT-PCR, Western Blot, Over Expression

Journal: Frontiers in Oncology

Article Title: COL8A1 Promotes NSCLC Progression Through IFIT1/IFIT3-Mediated EGFR Activation

doi: 10.3389/fonc.2022.707525

Figure Lengend Snippet: IFIT1 and IFIT3 knockdown effectively suppresses proliferation, migration, and apoptosis of NSCLC cells. (A, B) Effects of IFIT1 and IFIT3 knockdown on EGFR and AKT activity determined by western blotting. (C, D) Cell viability assays performed in IFIT1-knockdown H1299 and H1975 cells. (E, F) Cell viability assays performed in IFIT3-knockdown H1299 and H1975 cells. (G, H) Colony formation assessed in siCtrl-, siIFIT1-1, and siIFIT3-1-treated H1299 (G) and H1975 (H) cells. (I) Effects of IFIT1 and IFIT3 knockdown on apoptosis assessed by flow cytometry. (J) Effects of IFIT1 and IFIT3 knockdown on cell invasion and migration assessed by transwell assay. (K, L) H1299 and H1975 cells transfected with COL8A1, siIFIT1, and siIFIT3 were subjected to cell viability (K) and colony formation analysis (L) . **p < 0.01. ***p < 0.001.

Article Snippet: Primary antibodies against p-EGFR (#3777S, CST), EGFR (#2085, CST), p-AKT (#4060, CST), AKT (#4821, CST), IFIT-1 (23247-1-AP, Proteintech), IFIT-3 (15201-1-AP, Proteintech), and β-actin (#4970, CST) were applied in this study.

Techniques: Knockdown, Migration, Activity Assay, Western Blot, Flow Cytometry, Transwell Assay, Transfection

Journal: Frontiers in Oncology

Article Title: COL8A1 Promotes NSCLC Progression Through IFIT1/IFIT3-Mediated EGFR Activation

doi: 10.3389/fonc.2022.707525

Figure Lengend Snippet: COL8A1 contributes to NSCLC progression by activating EGFR in vivo . BALB/c nude mice were subcutaneously transplanted with COL8A1-silenced NSCLC cells. Mice were sacrificed 5 weeks after implantation. (A) Macroscopic images of xenografted tumors. (B) Tumor growth curves. (C) Tumor weight was determined immediately after sacrificing the mice. (D) Tumor tissues were subjected to western blotting analysis with the indicated antibodies. ***p < 0.001.

Article Snippet: Primary antibodies against p-EGFR (#3777S, CST), EGFR (#2085, CST), p-AKT (#4060, CST), AKT (#4821, CST), IFIT-1 (23247-1-AP, Proteintech), IFIT-3 (15201-1-AP, Proteintech), and β-actin (#4970, CST) were applied in this study.

Techniques: In Vivo, Western Blot

Journal: Frontiers in Oncology

Article Title: COL8A1 Promotes NSCLC Progression Through IFIT1/IFIT3-Mediated EGFR Activation

doi: 10.3389/fonc.2022.707525

Figure Lengend Snippet: Analysis of COL8A1, IFIT1, IFIT3, and p-EGFR in NSCLC tissues. The expression of COL8A1, IFIT1, IFIT3, and the phosphorylation of EGFR in NSCLC tissues detected by IHC staining.

Article Snippet: Primary antibodies against p-EGFR (#3777S, CST), EGFR (#2085, CST), p-AKT (#4060, CST), AKT (#4821, CST), IFIT-1 (23247-1-AP, Proteintech), IFIT-3 (15201-1-AP, Proteintech), and β-actin (#4970, CST) were applied in this study.

Techniques: Expressing, Phospho-proteomics, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: COL8A1 Promotes NSCLC Progression Through IFIT1/IFIT3-Mediated EGFR Activation

doi: 10.3389/fonc.2022.707525

Figure Lengend Snippet: Spearman correlation analysis of expression among COL8A1, IFIT1, IFIT3 and p-EGFR in 50 NSCLC tissues by IHC.

Article Snippet: Primary antibodies against p-EGFR (#3777S, CST), EGFR (#2085, CST), p-AKT (#4060, CST), AKT (#4821, CST), IFIT-1 (23247-1-AP, Proteintech), IFIT-3 (15201-1-AP, Proteintech), and β-actin (#4970, CST) were applied in this study.

Techniques: Expressing

Journal: bioRxiv

Article Title: Betrixaban Activates cGAS and ERVs to Promote Dual Nucleic-Sensing Antiviral Immunity

doi: 10.1101/2025.08.08.669242

Figure Lengend Snippet: (A) Distribution of transposable element (TE) classes among loci up-regulated by BT in bulk RNA-seq (counts of LTR, DNA, LINE, SINE, Satellite and Unknown elements). (B) Reactome terms enriched among BT-reactivated ERV loci. (C) GSEA enrichment plots in BT-versus DMSO-treated HT1080 cells. (D) Confocal micrographs of RAW 264.7 cells treated with DMSO or 50 µM BT for 12 h, stained for dsRNA (green) and nuclei (DAPI, blue). BT treatment induces punctate cytosolic dsRNA accumulations. Scale bars, 5 µm. (E) RT-qPCR of IFNB1 mRNA in HeLa wild-type (WT), MDA5 knockout, RIG-I knockout and MAVS knockout cells treated with BT. (F) Secreted type I IFN measured by ELISA in the same HeLa cell lines and treatments as in (E). (G) Western blot of p-TBK1, total TBK1, MDA5, MAVS and RIG-I, the samples were same to (E). (H) Flow cytometric quantification of VSV-GFP infection in HeLa wild-type (WT), MDA5 knockout, RIG-I knockout and MAVS knockout cells treated with BT. (I-J) Left are survival curves of MAVS knockout C57BL/6J mice challenged with lethal VSV and HSV-1 after a single i.p. dose of BT (50 mg/kg). Right are viruses RNA levels in livers measured by RT-qPCR. RIP-qPCR detection of HERVK14-int RNA associated with RIG-I or MDA5 in RAW 264.7 cells treated with DMSO or BT for 12 h. (L) ELISA quantification of secreted type I IFN and RT-qPCR of IFNB1 mRNA. (M) Flow cytometry of VSV-GFP infection. Data are shown as mean ± SEM. N.S., not significant, p > 0.05; *p < 0.05; **p < 0.01; ****p < 0.0001.

Article Snippet: Rabbit antibodies against TBK1 (catalog no. 28397-1-AP), Beta Tubulin (catalog no. 10094-1-AP), RIG-1/DDX58 (catalog no. 25068-1-AP), MAVS (catalog no. 14341-1-AP) and IFIT3 (catalog no. 15201-1-AP), along with mouse antibodies targeting Beta Actin (catalog no. 66009-1-Ig) and the GFP tag (catalog no. 50430-2-AP), were procured from Proteintech.

Techniques: RNA Sequencing, Staining, Quantitative RT-PCR, Knock-Out, Enzyme-linked Immunosorbent Assay, Western Blot, Infection, Flow Cytometry