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. 2023 Nov 22;97(12):e01183-23. doi: 10.1128/jvi.01183-23

Fig 2.

Fig 2

NF-κB activates JINR1 expression during JEV infection. (A) JINR1 induction upon JEV infection plateaus. SH-SY5Y cells were infected with JEV (MOI 5), and JINR1 levels were measured at the indicated time points using qRT-PCR. (B) JINR1 induction upon JEV infection is MOI dependent. SH-SY5Y cells were infected with different MOI as indicated, and the JINR1 levels were measured by qRT-PCR at 48 hpi. (C) JINR1 induction upon DENV infection is time dependent. SH-SY5Y cells were infected with DENV (MOI 5), and the JINR1 levels were measured at the indicated time points using qRT-PCR. (D) JINR1 induction upon WNV infection is time dependent. SH-SY5Y cells were infected with WNV (MOI 5), and the JINR1 levels were measured at the indicated time points using qRT-PCR. (E) Poly (I:C) induces JINR1 expression. SH-SY5Y cells were transfected with different amounts of poly(I:C), as indicated for 24 h. JINR1 transcript levels were determined by qRT-PCR. (F) JEV promotes RNA Pol II and H3K4me3 recruitment at the promoter of JINR1. Relative enrichment of RNA Pol II and H3K4me3 at the JINR1 promoter region in MI or JEV-infected SH-SY5Y cells determined by ChIP-qRT-PCR 36 hpi. Enrichment values are relative to MI-IgG. (G) NF-κB inhibition abrogates JINR1 induction upon JEV infection. SH-SY5Y and T98G cells were pre-treated with 1 µM of BAY11-7085 for 2 h, followed by JEV infection (MOI 5). JINR1 transcript levels were determined by qRT-PCR at 48 hpi. (H) JEV promotes the p65 subunit of NF-κB recruitment at the promoter of JINR1. Relative enrichment of p65 at the JINR1 promoter in MI or JEV-infected SH-SY5Y cells determined by ChIP-qRT-PCR 36 hpi. Enrichment values are relative to MI-IgG. Error bars represent the mean ± SEM from three independent experiments. Statistical comparisons were made using Student’s t-test. (A–E and G) RNA samples were analyzed by qRT-PCR. (A–D) *Significant change compared to the MI. (E) *Significant change compared to the control. (F and H) ChIP purified DNA was analyzed by qRT-PCR, and error bars represent the mean ± SEM from three independent experiments. *Significant change compared to the MI. Statistical comparisons were made using Student’s t-test. (G) *Significant change compared to the respective MI sample. #Significant change from JEV-infected sample.