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. 2023 Jul 7;14:1177324. doi: 10.3389/fimmu.2023.1177324

Figure 2.

Figure 2

In vitro characterization of IAV-NS1. (A) Schematic representation of the NA-TBEV NS1 gene segment, NCR = non-coding region, ATG = start codon, *Stop = stop codon. (B) PCR products specific for IAV NA performed on I = rPR8, II = IAV-RBD, III = IAV-NS1 and IV = negative control (1% agarose TBE gel, M = 1 kb ladder). (C) Immunostaining of rPR8, IAV-RBD or IAV-NS1 infected MDCK cells (MOI 0.01). 24 hpi, cells were fixed with 4% PFA, for intracellular staining permeabilized with TritonX®-100 and immunostained against IAV NP or TBEV NS1 (10x magnification). (D) Western blot analysis of whole cell lysate from MDCK cells infected with rPR8, IAV-RBD or IAV-NS1 for 24 h (MOI 0.01). Blots were stained against TBEV NS1. For control, antibodies against IAV nucleoprotein (NP) and GAPDH were included. (E) Growth curves for rPR8 (black), IAV-RBD (gray) and IAV-NS1 (red) in the presence (solid line) or absence (dotted line) of eNA on MDCK cells (MOI 0.001). Mann-Whitney test was used for statistical comparison between rPR8 –eNA and IAV-RBD –eNA or IAV-NS1 –eNA, respectively (* p<0.05).