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. 2022 Dec 1;13:1071394. doi: 10.3389/fmicb.2022.1071394

Figure 7.

Figure 7

miR-615 inhibits NF-κB pathway activation by regulating IRAK1. (A) IRAK1-flag or negative control (NC) and (B) si-IRAK1 or siRNA control (SC) were co-transfected with pNiFty-luc and pRL-TK into IECs. Cells were then stimulated with poly (I:C) and infected with PEDV (MOI = 1) at 24 h post-infection (hpi) or left untreated (unstimulated). Cells were then collected for the dual-luciferase reporter assay. (C,D) IECs were transfected with IRAK1-flag (or NC) or si-IRAK1 (or SC) for 24 h. Poly (I:C) stimulation and viral infection were then induced. After 24 h, IECs were collected for RT-qPCR to determine (C) IFN-λ1 and (D) IFN-λ3 expression. (E) Rescue experiment. Luciferase activity was rescued by IRAK1 overexpression. IECs were co-transfected with miR-615 (or MC) and IRAK1-flag (or NC). At 24 hpi, poly (I:C) stimulation and viral infection were induced. After 24 h of stimulation, cells were collected and the luminescence activity was determined. (F) IRAK1 knockdown and overexpression were confirmed using western blotting. The data are representative of three independent experiments (mean ± SD). ***p < 0.001.