polyclonal rabbit anti irf7  (Boster Bio)

Journal: Journal of Virology

Article Title: Cellular transcription factor TFII-I represses adenovirus gene expression

doi: 10.1128/jvi.00618-25

Figure Lengend Snippet: TFII-I KO does not alter the DDR or IFN response to Ad5 infection. TFII-I KO cells were generated by infecting parental BEC, NBS1-, or HDF cells with a CRISPR-Cas9 and guide RNA containing Lentivirus, followed by clonal selection. TFII-I KO was confirmed via Western blot (A, BEC; C, NBS1-; and G, HDF); α-tubulin or vimentin was measured as loading controls. Two independent TFII-I KO clones were selected for each cell type. ( B ) Parental and TFII-I KO BECs were infected with E4ORF3 - /E4ORF6 - Ad5 at an MOI of 1, and total cellular DNA was isolated at the indicated times post-infection. HAdV genome levels were quantified by qPCR and normalized to endogenous GAPDH levels. The results shown represent the mean values ± SD; n = 3. ( D ) NBS1 - (NBS/vector), NBS1 - ectopically expressing Nbs1 (NBS/Nbs1), and NBS1 - TFII-I KO cells (NBS TFII-I-/- #8 and #9) were infected with E4ORF3 - /E4ORF6 - Ad5 (dl355/inORF3) at an MOI of 1, and total cellular DNA was isolated at the indicated time post-infection. HAdV genome levels were quantified by qPCR and normalized to endogenous GAPDH levels. The results shown represent the mean values ± SD; n = 3. ( E ) Parental and TFII-I KO BECs were pretreated for 24 h with IFNα. IRF7 levels were measured by Western blot; Vimentin was measured as a loading control. ( F ) Parental and TFII-I KO HDFs were pretreated for 24 h with IFN-α or IFN-γ and infected with Ad5-WT at an MOI of 1. Total cellular DNA was isolated at the indicated times post-infection. HAdV genome levels were quantified by qPCR and normalized to endogenous GAPDH levels. The results shown represent the mean values ± SD; n = 3.

Article Snippet: The antibodies used were as follows: rabbit polyclonal TFII-I antibody (Cell Signaling Technologies) (1:1,000 dilution), rabbit polyclonal IRF7 (Cell Signaling Technologies) (1:1,000), mouse monoclonal hexon antibody (MAB0850; Abnova) (1:5,000 dilution), rabbit polyclonal penton antibody (gift from Carl Anderson, Brookhaven National Laboratory) (1:1,000 dilution), rabbit polyclonal IVa2 antibody ( ) (1:1,000 dilution), rabbit polyclonal L1-52/55K antibody ( ) (1:1,000 dilution), rabbit polyclonal V antibody (gift from David Matthews, University of Bristol) (1:1,000 dilution), rabbit polyclonal L4-100K antibody ( ) (1:1,000 dilution), rabbit polyclonal VIII antibody (gift from Ann Tollefson and William Wold, St. Louis University) (1:400 dilution), rabbit polyclonal E1A antibody (SC430; Santa Cruz Biotechnology) (1:500 dilution), rabbit polyclonal DNA binding protein (DBP) antibody (gift from Peter van der Vleit, University Utrecht, Netherlands) (1:1,000 dilution), mouse monoclonal α-Tubulin (Sigma-Millipore #T5168) (1:10,000), rabbit polyclonal GAPDH (Millipore Sigma Aldrich) (1:7,500), and rabbit monoclonal vimentin (NeoMarkers) (1:2,000).

Techniques: Infection, Generated, CRISPR, Selection, Western Blot, Clone Assay, Isolation, Plasmid Preparation, Expressing, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Genome-wide analysis reveals the MORC3-mediated repression of PD-L1 expression in head and neck cancer

doi: 10.3389/fcell.2024.1410130

Figure Lengend Snippet: MORC3 represses the expression of many IFN-associated target genes. (A) The expression levels of IFN-associated target genes MX2, IFIT1, IRF7, IRF9, IFITM3, IFITM1, IFI44, and IFIH1 in CAL 27 cells with or without MORC3 knockdown were analyzed by qRT-PCR. (B) The expression levels of IFN-associated target genes MX2, IFIT1, IRF7, IRF9, IFITM3, IFITM1, and IFI44 in CAL 27 cells with or without MORC3 overexpression were analyzed by qRT-PCR. (C) The correlation between the expression levels of MORC3 and IFN-associated target genes in single cells of head and neck cancer were evaluated by reanalyzing the expression data of GSE10332 in the database of CancerSEA. *, p < 0.05.

Article Snippet: The membranes were blocked with 5% skimmed milk and incubated overnight at 4°C with following antibodies: mouse monoclonal anti-MORC3 antibody (Santa Cruz Biotechnology, United States), rabbit monoclonal anti- interferon induced protein with tetratricopeptide repeats 2 (IFIT2) antibody (abcam, United States), rabbit monoclonal anti-interferon induced transmembrane protein 3 (IFITM3) antibody (Cell Signaling Technology, United States), rabbit monoclonal anti-interferon induced protein with tetratricopeptide repeats 1 (IFIT1) antibody (Cell Signaling Technology, United States), rabbit monoclonal anti-cyclin D1 (CCND1) (ABclonal, China), rabbit monoclonal anti-cyclinD2 (CCND2) (ABclonal, China), rabbit polyclonal anti-JUN (ABclonal, China), rabbit polyclonal anti-interferon regulatory factor 7 (IRF7) (Proteintech, China), rabbit polyclonal anti-PD-L1 antibody (Proteintech, China), rabbit polyclonal anti-DExD/H-Box Helicase 60 (DDX60) antibody (Proteintech, China), and mouse anti-β-actin (Abmart, China).

Techniques: Expressing, Knockdown, Quantitative RT-PCR, Over Expression