chicken embryo fibroblast df 1 cells  (ATCC)

Journal: bioRxiv

Article Title: RNA length and receptor usage define innate immune recognition across species

doi: 10.64898/2026.05.21.726451

Figure Lengend Snippet: Induction of antiviral response in human (A549), chicken (DF-1), and duck (CCL-141) cells following transfection with an in vitro transcribed mvRNA. The mvRNA used in this study is a 60 nt RNA forming an approximately 28 bp duplex and bearing a 5’-triphosphate (5’-ppp) or dephosphorylated (5’-OH) end. Luciferase activity driven by interferon-responsive promoters is shown. Cells were transfected with mvRNA for 24 h prior to measurement. Bars represent mean ± SEM from three independent biological replicates (each with three technical replicates), and individual points indicate technical replicates. Statistical significance: ****P < 0.0001; ns, not significant (one-way ANOVA with Tukey’s multiple comparisons test).

Article Snippet: Human lung epithelial A549 cells (CCL-185) and chicken embryo fibroblast DF-1 cells (CRL-3586) were obtained from ATCC.

Techniques: Transfection, In Vitro, Luciferase, Activity Assay

Journal: bioRxiv

Article Title: RNA length and receptor usage define innate immune recognition across species

doi: 10.64898/2026.05.21.726451

Figure Lengend Snippet: (A) Induction of antiviral response in human (A549), chicken (DF-1), and duck (CCL-141) cells following transfection with in vitro transcribed dsRNAs of defined lengths (50, 200, 550, and 1600 bp). dsRNAs carried either a 5’-triphosphate (5’-ppp) or a dephosphorylated (5’-OH) end. Luciferase activity driven by interferon-responsive promoters is shown. Cells were transfected with the indicated dsRNA for 24 h prior to measurement. Bars represent mean ± SEM from three independent biological replicates (each with three technical replicates), and individual points indicate technical replicates. Statistical significance is shown relative to mock-treated samples: **P < 0.01; ****P < 0.0001; ns, not significant (one-way ANOVA with Tukey’s multiple comparisons test). (B) Western blot analysis of MX1 protein expression in human and chicken cells following transfection with mvRNA and 5’-triphosphorylated dsRNAs of increasing lengths (50-550 bp).

Article Snippet: Human lung epithelial A549 cells (CCL-185) and chicken embryo fibroblast DF-1 cells (CRL-3586) were obtained from ATCC.

Techniques: Transfection, In Vitro, Luciferase, Activity Assay, Western Blot, Expressing

Journal: bioRxiv

Article Title: RNA length and receptor usage define innate immune recognition across species

doi: 10.64898/2026.05.21.726451

Figure Lengend Snippet: (A) Schematic overview of RNA fractionation from virus (H5N8 or H1N1)-infected, dsRNA-treated, and mock-treated A549 cells using HPLC. Total RNA was fractionated, and RNA species shorter than 200 nt were collected. The short RNA fraction, either untreated or dephosphorylated with alkaline phosphatase, was used for transfection of human (A549) and chicken (DF-1) cells. (B) Northern blot analysis confirming the presence of endogenous short RNA, SNORD44 (63 nt), in the collected fraction. (C) Induction of antiviral response in human and chicken cells following transfection with the isolated short RNA fraction, assessed by western blot analysis of MX1 protein expression. (D) Analysis of total RNA isolated from virus-infected, dsRNA-treated, and mock-treated A549 cells. rRNA integrity was assessed in a denaturing agarose gel.

Article Snippet: Human lung epithelial A549 cells (CCL-185) and chicken embryo fibroblast DF-1 cells (CRL-3586) were obtained from ATCC.

Techniques: Fractionation, Virus, Infection, Transfection, Northern Blot, Isolation, Western Blot, Expressing, Agarose Gel Electrophoresis

Journal: bioRxiv

Article Title: RNA length and receptor usage define innate immune recognition across species

doi: 10.64898/2026.05.21.726451

Figure Lengend Snippet: (A) Western blot analysis of antiviral responses assessed by MX1 expression in human (A549) cells lacking RIG-I or MDA5 following transfection with 5’-triphosphorylated in vitro transcribed mvRNA and 5’-triphosphorylated dsRNAs of increasing lengths. Lanes labeled “A549 WT” represent wild-type A549 cells transfected with 5’-ppp mvRNA and are included as positive controls for immune activation. (B) Induction of antiviral response in human cells lacking RIG-I or MDA5 following transfection with the isolated short RNA fraction, assessed by western blot analysis of MX1 expression. Lanes labeled “A549 WT” represent wild-type A549 cells transfected with with 5’-ppp mvRNA and are included as positive controls for immune activation. (C) Induction of antiviral response in chicken cells lacking MDA5 following transfection with 5’-triphosphorylated in vitro transcribed mvRNA and 5’-triphosphorylated dsRNAs of increasing lengths, assessed by western blot analysis of MX1 expression. Lanes labeled “DF-1 WT” represent wild-type DF-1 cells transfected with 5’-ppp 550 bp dsRNA and are included as positive controls for immune activation.

Article Snippet: Human lung epithelial A549 cells (CCL-185) and chicken embryo fibroblast DF-1 cells (CRL-3586) were obtained from ATCC.

Techniques: Western Blot, Expressing, Transfection, In Vitro, Labeling, Activation Assay, Isolation

Journal: bioRxiv

Article Title: RNA length and receptor usage define innate immune recognition across species

doi: 10.64898/2026.05.21.726451

Figure Lengend Snippet: (A) Western blot analysis of PKR and eIF2α phosphorylation in human (A549), chicken (DF-1), and duck (CCL-141) cells following transfection with short (mvRNA) or long (1600 bp) dsRNA carrying either a 5’-triphosphate (5’-ppp) or a dephosphorylated (5’-OH) end. Human and chicken cells were transfected for 5 h, whereas duck cells were transfected for 7.5 h prior to harvesting. PKR phosphorylation was assessed in human samples. (B) Puromycin incorporation assay to assess global translation in human, chicken, and duck cells following transfection with short or long dsRNA. Reduced puromycin signal indicates translational inhibition.

Article Snippet: Human lung epithelial A549 cells (CCL-185) and chicken embryo fibroblast DF-1 cells (CRL-3586) were obtained from ATCC.

Techniques: Western Blot, Phospho-proteomics, Transfection, Inhibition

Journal: bioRxiv

Article Title: RNA length and receptor usage define innate immune recognition across species

doi: 10.64898/2026.05.21.726451

Figure Lengend Snippet: (A) Schematic overview of the pull-down assay. Human (A549), chicken (DF-1), and duck (CCL-141) cells were transfected with a mixture of 5’-triphosphorylated short (mvRNA) and long (1600 bp) dsRNA or mock-treated prior to lysis. Cell lysates were incubated with biotinylated short or long dsRNA carrying either a 5’-triphosphate (5’-ppp) or a dephosphorylated (5’-OH) end. RNA-protein complexes were isolated using streptavidin beads and analyzed by mass spectrometry (MS/MS). (B) Heatmaps showing enrichment of RNA-binding proteins in pull-down assays from human, chicken, and duck cell lysates. Protein enrichment is displayed as a composite score calculated as log2(FC) multiplied by -log10(p-value), relative to beads-only control samples. Color scales indicate relative protein enrichment. The top 30 enriched proteins are shown; full datasets are provided in the supporting information (Fig. S11).

Article Snippet: Human lung epithelial A549 cells (CCL-185) and chicken embryo fibroblast DF-1 cells (CRL-3586) were obtained from ATCC.

Techniques: Pull Down Assay, Transfection, Lysis, Incubation, Isolation, Mass Spectrometry, Tandem Mass Spectroscopy, RNA Binding Assay, Protein Enrichment, Control

Journal: bioRxiv

Article Title: RNA length and receptor usage define innate immune recognition across species

doi: 10.64898/2026.05.21.726451

Figure Lengend Snippet: Analysis of total RNA integrity in human (A549), chicken (DF-1), and duck (CCL-141) cells following transfection with short (mvRNA) or long (1600 bp) dsRNA carrying either a 5’-triphosphate (5’-ppp) or a dephosphorylated (5’-OH) end. RNA profiles were assessed using Agilent 2100 Bioanalyzer.

Article Snippet: Human lung epithelial A549 cells (CCL-185) and chicken embryo fibroblast DF-1 cells (CRL-3586) were obtained from ATCC.

Techniques: Transfection

Journal: Poultry Science

Article Title: Highly efficient gene editing via targeted Cas9 insertion into chicken housekeeping gene

doi: 10.1016/j.psj.2026.106585

Figure Lengend Snippet: CRISPR/Cas9-mediated targeting of ACTB and GAPDH genes in chicken DF-1 cells. (A, F) Schematic diagrams of the ACTB (A) and GAPDH (F) gene structures, showing CRISPR/Cas9 targeting sites. (B–E) Validation of ACTB targeting vectors. (B, D) T7E1 assays and (C, E) Sanger sequencing of DF-1 cells transfected with CRISPR/Cas9 constructs targeting the 3′ region (B, C) or intron (D, E). (G–J) Validation of GAPDH targeting vectors. (G, I) T7E1 assays and (H, J) Sanger sequencing of DF-1 cells transfected with constructs targeting the 3′ region (G, H) or intron (I, J). gRNA sequences are shown in red or blue, PAM sequences in yellow. Deleted bases are indicated by strikethrough lines, substitutions by italics, and insertions by lowercase letters.

Article Snippet: Chicken DF-1 fibroblast cells (ATCC® CRL-12203, American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Cytiva, Marlborough, MA, USA) and 1 × antibiotic-antimycotic solution (Gibco).

Techniques: CRISPR, Biomarker Discovery, Sequencing, Transfection, Construct

Journal: Poultry Science

Article Title: Highly efficient gene editing via targeted Cas9 insertion into chicken housekeeping gene

doi: 10.1016/j.psj.2026.106585

Figure Lengend Snippet: Validation of Cas9-GFP knock-in at the ACTB and GAPDH loci in DF-1 Cells. (A) Schematic illustration of the 3′ region targeted and tagging CRISPR/Cas9 approaches. (B) Detection of GFP in ACTB and GAPDH targeted chicken DF-1 cells. Non-transfected wild-type (WT) DF-1 cells are shown as a control, appearing without fluorescence under standard and fluorescence microscopy. Cells successfully transfected with the knock-in vector constructs targeting ACTB and GAPDH genes exhibit green fluorescence, indicating expression of the reporter gene. Scale bar, 100 µm. (C) Knock-in-specific junction PCR of targeted sites. (D, F) Sequencing analysis of the 3′ region targeted knock-in in chicken DF-1 cells. The schematic illustrates the gene locus following CRISPR/Cas9-mediated insertion of a donor cassette at the 3′ region targeting site via non-homologous end joining (NHEJ). Sanger sequencing of the junction PCR products confirmed integration of the donor sequence in the adjacent genomic regions with indel mutations. (E, G) This schematic depicts the post-integration structure of each gene following CRISPR/Cas9-NHEJ-mediated targeted gene tagging. The donor plasmid was designed with GFP flanked by genomic homology arms corresponding to sequences adjacent to the targeted intron. Sanger sequencing of the junction PCR products confirmed integration of the donor sequence in the adjacent genomic regions with indel mutation.

Article Snippet: Chicken DF-1 fibroblast cells (ATCC® CRL-12203, American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Cytiva, Marlborough, MA, USA) and 1 × antibiotic-antimycotic solution (Gibco).

Techniques: Biomarker Discovery, Knock-In, CRISPR, Transfection, Control, Fluorescence, Microscopy, Plasmid Preparation, Construct, Expressing, Sequencing, Non-Homologous End Joining, Mutagenesis

Journal: Poultry Science

Article Title: Highly efficient gene editing via targeted Cas9 insertion into chicken housekeeping gene

doi: 10.1016/j.psj.2026.106585

Figure Lengend Snippet: Validation of Cas9 activity in ACTB and GAPDH knock-in (KI) chicken DF-1 cells. (A) Gene structure of the intergenic region between DMRT1 and DMRT3 is depicted, showing exons as boxes and introns as lines, with the gRNA target site indicated. (B) T7E1 assay for KI DF-1 cells ( ACTB 3′ KI, ACTB tagging, GAPDH 3′ KI, and GAPDH tagging) followed by transfection with gRNA expressing vector. (C) Sanger sequencing analysis of KI chicken DF-1 cells ( GAPDH 3′ KI, and GAPDH tagging) transfected with DMRT gRNA are shown. gRNA sequences are shown in red, PAM sequences in yellow. The strikethrough lines indicate regions where base pairs have been deleted.

Article Snippet: Chicken DF-1 fibroblast cells (ATCC® CRL-12203, American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Cytiva, Marlborough, MA, USA) and 1 × antibiotic-antimycotic solution (Gibco).

Techniques: Biomarker Discovery, Activity Assay, Knock-In, Transfection, Expressing, Plasmid Preparation, Sequencing

Journal: Poultry Science

Article Title: Highly efficient gene editing via targeted Cas9 insertion into chicken housekeeping gene

doi: 10.1016/j.psj.2026.106585

Figure Lengend Snippet: Generation and validation of single-cell clones with Cas9-GFP knock-in at the GAPDH locus in chicken DF-1 cells. (A) Bright-field (BF) and GFP fluorescence images obtained after subculture following single-cell seeding. Each panel represents a clonal population derived from a single genome-edited cell. A total of 16 single-cell-derived clones were identified from the 96-well plates, of which 12 maintained consistent growth after subculture. Clone numbers correspond to the original 16 identified clones, and images of the 12 viable clones are shown. Scale bar, 100 µm. (B) PCR analysis of 12 single-cell-derived clones following subculture. Intron-targeted knock-in alleles were confirmed by 5′ junction PCR using junction-specific primers. The presence of residual wild-type (WT) alleles in individual clones was assessed using WT allele–specific primers. GAPDH PCR served as a genomic DNA quality control. (C) Relative Cas9 copy number was estimated by quantitative PCR (qPCR) using genomic DNA from each clone, normalized to the endogenous GAPDH reference locus (two copies in diploid cells). Bars represent the mean ± SD of technical qPCR replicates ( n = 3).

Article Snippet: Chicken DF-1 fibroblast cells (ATCC® CRL-12203, American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Cytiva, Marlborough, MA, USA) and 1 × antibiotic-antimycotic solution (Gibco).

Techniques: Biomarker Discovery, Single Cell, Clone Assay, Knock-In, Fluorescence, Derivative Assay, Control, Real-time Polymerase Chain Reaction

Journal: Poultry Science

Article Title: Highly efficient gene editing via targeted Cas9 insertion into chicken housekeeping gene

doi: 10.1016/j.psj.2026.106585

Figure Lengend Snippet: Characterization of single-cell-derived Cas9-expressing DF-1 clones. (A) Flow cytometry analysis of GFP expression levels in GAPDH tagging clones. (B) Median fluorescence intensity (MFI) of GFP in each clone. Data represents n = 3 biological replicates; bars show mean ± SD. ⁎⁎⁎⁎ P < 0.0001. (C) Western blot analysis of Cas9 and GAPDH protein expression in each clone. α-tubulin was used as a loading control. (D–E) Functional validation of genome editing capability in single-cell-derived Cas9-expressing DF-1 clones. A guide RNA (gRNA) expression vector targeting an internal region between DMRT1 and DMRT3 was transfected into each clone. As a control, wild-type (WT) DF-1 cells were co-transfected with the same gRNA vector and a transient Cas9 expression plasmid. (D) Genome editing activity was assessed by T7 endonuclease I (T7E1) assay. (E) Sanger sequencing of the target site confirmed indel formation at the expected genomic locus. gRNA sequences are shown in red, PAM sequences in yellow. Deleted bases are indicated by strikethrough lines, substitutions by italics, and insertions by lowercase letters.

Article Snippet: Chicken DF-1 fibroblast cells (ATCC® CRL-12203, American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; HyClone, Cytiva, Marlborough, MA, USA) and 1 × antibiotic-antimycotic solution (Gibco).

Techniques: Single Cell, Derivative Assay, Expressing, Clone Assay, Flow Cytometry, Fluorescence, Western Blot, Control, Functional Assay, Biomarker Discovery, Plasmid Preparation, Transfection, Activity Assay, Sequencing

Journal: Frontiers in Immunology

Article Title: Chicken IRF10 suppresses the cGAS-STING-IFN antiviral signaling pathway by targeting IRF7

doi: 10.3389/fimmu.2026.1767491

Figure Lengend Snippet: Analysis of chicken (ch) IRF family members in cGAS-STING-IFN signaling by promoter assays. (A) Flag-tagged chIRF family members were co-transfected with chGAS-STING into 293T cells, and ISRE promoter activity was measured at 24 h post-transfection. (B) Flag-tagged chIRF family members were co-transfected with chSTING into DF-1 cells, and chIFN-β promoter activity was measured at 24 h post-transfection. (C) GFP-tagged chIRF family members were co-transfected with chSTING into DF-1 cells, and chIFN-β promoter activity was measured at 24 h post-transfection. ** p < 0.01 versus vector controls.

Article Snippet: The HEK-293T cells (ATCC Cat# CRL-3216) and chicken fibroblast DF-1 cells (ATCC Cat# CRL-12203) were maintained in DMEM (Hyclone Laboratories, USA) supplemented with 10% fetal bovine serum (FBS, Vazyme Biotech).

Techniques: Transfection, Activity Assay, Plasmid Preparation

Journal: Frontiers in Immunology

Article Title: Chicken IRF10 suppresses the cGAS-STING-IFN antiviral signaling pathway by targeting IRF7

doi: 10.3389/fimmu.2026.1767491

Figure Lengend Snippet: chIRF10 negatively regulates the chicken cGAS-STING-IFN signaling. (A, B) HD11 cells were transfected with increasing doses of chIRF10, which was normalized with pCMV vector. At 12 h post-transfection, the cells were stimulated with 2 μg/mL cGAMP (A) or 1 μg/mL poly dA:dT (B) for 12 h, followed by measurement of chIFN-β promoter activity. (C) DF-1 cells were co-transfected with chSTING and increasing doses of chIRF10, normalized with pCMV vector. Cells were collected at 24 h post-transfection to measure chIFN-β promoter activity. (D – F) HD11 cells were transfected with either chIRF10 or pCMV vector. At 12 h post-transfection, the cells were stimulated with cGAMP or poly dA:dT for 24 h, and the mRNA expression levels of downstream genes IFN-β (D) , MX1 (E) , and OASL (F) were detected by RT-qPCR. (G – J) )DF-1 cells were co-transfected with chSTING and increasing doses of chIRF10 for 48 h, and the mRNA expression levels of downstream genes IFN-β (G) , MX1 (H) , OASL (I) , and PKR (J) were measured by RT-qPCR. * p < 0.05 and ** p < 0.01 versus vector controls.

Article Snippet: The HEK-293T cells (ATCC Cat# CRL-3216) and chicken fibroblast DF-1 cells (ATCC Cat# CRL-12203) were maintained in DMEM (Hyclone Laboratories, USA) supplemented with 10% fetal bovine serum (FBS, Vazyme Biotech).

Techniques: Transfection, Plasmid Preparation, Activity Assay, Expressing, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Chicken IRF10 suppresses the cGAS-STING-IFN antiviral signaling pathway by targeting IRF7

doi: 10.3389/fimmu.2026.1767491

Figure Lengend Snippet: chIRF10 negatively regulates the antiviral function of chcGAS-STING signaling pathway. (A–D) HD11 cells were transfected with either chIRF10 or pCMV vector for 12 h, and then stimulated with cGAMP (A, B) or poly dA:dT (C, D) for 12 h, followed by infection with NDV-RFP at 0.01 MOI for 12 (h) RFP fluorescence was detected using fluorescence microscopy (A, C) , and cells were collected for WB analysis of RFP protein expression (B, D) . (E, F) HD11 cells were transfected and stimulated as in A, followed by infection with AIV (H1N1) at 0.01 MOI for 24 (h) The viral NP protein expression was detected by WB. (G–J) HD11 cells were transfected and stimulated as in A, followed by infection with SMV at 0.1 MOI for 24 h (G, H) or infection with VACV at 0.1 MOI for 24 h (I, J) . The viral copy number was measured by qPCR. (K, L) DF-1 cells were transfected with chIRF10 and chSTING as indicated for 24 h, and infected with NDV-RFP at 0.01 MOI for 12 h, followed by RFP fluorescence detection by fluorescence microscopy (K) and WB analysis of RFP protein expression (L–N) DF-1 cells were transfected as indicated for 24 h, and infected with SMV (M) or VACV (N) at 0.1 MOI for 24 h, followed by qPCR measurement of viral copy number. (O) HD11 cells were stimulated with cGAMP or infected with the corresponding viruses at the indicated doses. At 24 h post-infection, cells were collected and chIRF10 mRNA expression was detected by RT-qPCR. * p < 0.05 and ** p < 0.01 versus controls.

Article Snippet: The HEK-293T cells (ATCC Cat# CRL-3216) and chicken fibroblast DF-1 cells (ATCC Cat# CRL-12203) were maintained in DMEM (Hyclone Laboratories, USA) supplemented with 10% fetal bovine serum (FBS, Vazyme Biotech).

Techniques: Transfection, Plasmid Preparation, Infection, Fluorescence, Microscopy, Expressing, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Chicken IRF10 suppresses the cGAS-STING-IFN antiviral signaling pathway by targeting IRF7

doi: 10.3389/fimmu.2026.1767491

Figure Lengend Snippet: Deletion of the IAD, but not the DBD, abolishes the negative regulatory activity of chIRF10. (A) DF-1 cells were co-transfected with chIRF10 or its deletion mutants, with or without chSTING for 24 h, followed by measurement of chIFN-β promoter activity. (B–D) DF-1 cells were transfected as in A for 48 h, and the mRNA expressions of chSTING-activated downstream genes IFN-β (B) , MX1 (C) , and OASL (D) were detected by RT-qPCR. (E, F) DF-1 cells were transfected as indicated for 24 h and infected with NDV at 0.01 MOI for 12 (h) RFP fluorescence was observed using fluorescence microscopy (E) and RFP protein expression was detected by WB (F–H) DF-1 cells were transfected as indicated for 24 h and infected with SMV (G) or VACV (H) at 0.1 MOI for 24 (h) The viral copy number was measured by qPCR. ** p < 0.01 versus controls.

Article Snippet: The HEK-293T cells (ATCC Cat# CRL-3216) and chicken fibroblast DF-1 cells (ATCC Cat# CRL-12203) were maintained in DMEM (Hyclone Laboratories, USA) supplemented with 10% fetal bovine serum (FBS, Vazyme Biotech).

Techniques: Activity Assay, Transfection, Quantitative RT-PCR, Infection, Fluorescence, Microscopy, Expressing

Journal: Frontiers in Immunology

Article Title: Chicken IRF10 suppresses the cGAS-STING-IFN antiviral signaling pathway by targeting IRF7

doi: 10.3389/fimmu.2026.1767491

Figure Lengend Snippet: chIRF10 inhibits the IFN signaling activated by chTBK1, chIKKϵ, and chIRF7. (A) DF-1 cells were transfected with chIRF10 and chTBK1 as indicated. (B) DF-1 cells were transfected with chIRF10 and chIKKϵ as indicated. (C) DF-1 cells were transfected with chIRF10 and chIRF7 as indicated. chIFN-β promoter activity was measured at 24 h post-transfection, and the mRNA expression of downstream activated genes IFN-β, MX1, OASL, and PKR was detected by RT-qPCR at 48 h post-transfection. * p < 0.05 and ** p < 0.01 versus controls.

Article Snippet: The HEK-293T cells (ATCC Cat# CRL-3216) and chicken fibroblast DF-1 cells (ATCC Cat# CRL-12203) were maintained in DMEM (Hyclone Laboratories, USA) supplemented with 10% fetal bovine serum (FBS, Vazyme Biotech).

Techniques: Transfection, Activity Assay, Expressing, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Chicken IRF10 suppresses the cGAS-STING-IFN antiviral signaling pathway by targeting IRF7

doi: 10.3389/fimmu.2026.1767491

Figure Lengend Snippet: chIRF10 targets chIRF7 and inhibits chIRF7 activation. (A) DF-1 cells were transfected with chIRF10 and its deletion mutants, with or without chIRF7 as indicated for 24 (h) The co-localization of chIRF10/mutants and chIRF7 was examined by laser scanning confocal microscopy following IFA staining. (B, C) 293T cells were transfected with chIRF10 and chIRF7 (B) , chIRF10/chIRF10ΔDBD/chIRF10ΔIAD and chIRF7 (C) as indicated for 48 (h) The protein interactions between chIRF10/mutants and chIRF7 were assessed by co-IP using the indicated antibodies. (D – F) 293T cells were transfected with chIRF10, chIRF7 plus chcGAS-STING (D) , chIRF10/chIRF10ΔIAD, chIRF7 plus chcGAS-STING (E) , and increasing doses of chIRF10, chIRF7 plus chcGAS-STING (F) as indicated for 24 (h) The chIRF7 dimerization levels were analyzed by native PAGE. (G) 293T cells were co-transfected with GFP-tagged chIRF7, Flag-tagged chIRF10/chIRF10ΔIAD, and HA-tagged chcGAS-STING as indicated. The puncta formation of chIRF7 indicating its activation was observed by fluorescence microscopy 24 h post-transfection and marked as red arrows.

Article Snippet: The HEK-293T cells (ATCC Cat# CRL-3216) and chicken fibroblast DF-1 cells (ATCC Cat# CRL-12203) were maintained in DMEM (Hyclone Laboratories, USA) supplemented with 10% fetal bovine serum (FBS, Vazyme Biotech).

Techniques: Activation Assay, Transfection, Confocal Microscopy, Staining, Co-Immunoprecipitation Assay, Clear Native PAGE, Fluorescence, Microscopy