RBP4 promotes IAV replication <i>in vitro.</i>
<p>(A) Statistical analysis of the FPKM values of RBP4 in the blood of patients with influenza infection (n = 148) and healthy people (n = 82) obtained from four online GEO databases (GSE155635, GSE158592, GSE196350 and GSE213168). (B) qPCR analysis of RBP4 mRNA levels in A549, HEK293T and PK-...
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2025
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| Summary: | <p>(A) Statistical analysis of the FPKM values of RBP4 in the blood of patients with influenza infection (n = 148) and healthy people (n = 82) obtained from four online GEO databases (GSE155635, GSE158592, GSE196350 and GSE213168). (B) qPCR analysis of RBP4 mRNA levels in A549, HEK293T and PK-15 cells upon WSN (MOI = 0.1) infection at indicated periods. (C) ELISA of human RBP4 in supernatants of A549 and HEK293T cells infected with WSN (MOI = 0.1) for indicated periods. (D) Immunoblotting analysis of NP and M1 protein levels in HEK293T, PK-15 and 3D4/21 cells transfected with RBP4 or control plasmids for 12 hours, following infection with IAV WSN strain (MOI = 0.1) for various periods. (E, F) Immunoblotting (E) or qPCR analysis (F) of NP and M1 protein levels or mRNA expression in WT and RBP4-defecient PK-15 cells or HEK293T cells infected with IAV WSN strain (MOI = 0.1) for indicated periods. (G) Plaque assay of viral titers in WT and RBP4-deficient HEK293T and PK-15 cells infected with WSN (MOI = 0.1) for 24 hours. (H) Immunoblotting analysis of NP and M1 protein expression in WT and RBP4-deficient HEK293T cells infected with IAV PR8 strain (MOI = 0.1) for indicated periods. (I) Immunoblotting analysis of WT and RBP4-deficient PK-15 cells infected with IAV H3N2 (MOI = 0.1) or H9N2 (MOI = 0.1) for indicated periods. (J-L) RT-qPCR analysis of NP and M1 mRNA expression (J) or plaque assay of virus titers (L, left) in WT and RBP4-deficient HEK293T cells infected with IAV PR8 strain (MOI = 0.1) (J, L, left) or in WT and RBP4-deficient PK-15 cells infected with IAV H9N2 strain (K, left and L, middle) or H3N2 strain (K, right and L, right). (M) RT-qPCR analysis of mRNA levels of NP and M1 in RBP4-deficient PK-15 (RBP4-KO PK-15) cells infected with WSN (MOI = 0.1) for indicated periods. After 6 hours of WSN infection, cells were treated with or without recombinant porcine RBP4 (rpRBP4). Data are representative of two independent experiments (D, E, H, I) or pooled from at least three independent experiments (B, C, F, G, J-M, mean ± SD). <i><i>p</i></i> values were calculated using One-way ANOVA (B and C) or unpaired student’s <i><i>t</i></i>-test (A, F, G, J-M). *<i><i>p</i></i> < 0.05, **<i><i>p</i></i> < 0.01, ***<i><i>p</i></i> < 0.001, ****<i><i>p</i></i> < 0.0001.</p> |
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