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. 2017 Apr 13;91(9):e02250-16. doi: 10.1128/JVI.02250-16

FIG 3.

FIG 3

Alteration of NEAT1 affects HTNV-induced IFN expression in HUVECs. (A) HUVECs were infected with either 60Co-inactivated HTNV or HTNV at an MOI of 0.1 or 1. IFN-β expression at different time points was assessed by qRT-PCR (left) and ELISA (right). Values are means ± SD (n = 3; *, P < 0.01; Student's t test, compared with the 60Co-inactivated HTNV group). (B) HUVECs were infected with either 60Co-inactivated HTNV or HTNV at an MOI of 0.1 or 1. The NEAT1-2 expression at different time points was assessed by qRT-PCR. Values are means ± SD (n = 3; *, P < 0.01; Student's t test, compared with the 60Co-inactivated HTNV group). (C and D) HUVECs were transfected with NC or si-NEAT1-2 and, 24 h later, were infected with HTNV at an MOI of 0.1 (C) or 1 (D). IFN-β expression at different time points was assessed by qRT-PCR. Values are means ± SD (n = 3; *, P < 0.01; Student's t test, compared with the NC group). (E) HUVECs were transfected with control plasmids (vector) or pCMV-NEAT1-2 and, 24 h later, were infected with HTNV at an MOI of 1. IFN-β expression at different time points was assessed by qRT-PCR. Values are means ± SD (n = 3; *, P < 0.01; Student's t test, compared with the vector plus HTNV group). (F) Detection for luciferase reporter activities driven by the IFN-β promoter was done in siRNA or plasmid-pretransfected HUVECs without HTNV infection or with HTNV infection for 48 h (MOI = 5). Values are means ± SD (n = 5; *, P < 0.01; **, P < 0.001; Student's t test). (G) HUVECs were transfected with NC or si-NEAT1-2 and, 24 h later, were infected with HTNV at an MOI of 0.1. Increasing amounts of IFN-β were added starting at 12 hpi. The HTNV NP production was measured by Western blotting at 48 hpi. (H) HUVECs were transfected with control plasmids (vector) or pCMV-NEAT1-2 and, 24 h later, were infected with HTNV at an MOI of 1. Decreasing amounts of polyclonal sheep neutralizing antibodies against IFN-α and IFN-β (anti-IFN) were added starting at 12 hpi. The HTNV NP production was measured by Western blotting at 48 hpi.