Figure 5.

LPS inhibits CSFV replication in PAMs via the NF-κB-signalling pathway. (a) LPS treatment increases NF-κB activity in PAMs. PAMs were treated with LPS, and cell lysates were subjected to dual-luciferase reporter assays to determine NF-κB luciferase activity at 24 h and 48 h. (b,c) LPS treatment decreases CSFV replication in PAMs. RT-PCR analysed CSFV genome RNA, and viral titres in the supernatant were assessed and expressed as TCID₅₀/ml in LPS-treated PAMs at 24 and 48 hpi. (d) p65 knockdown by shRNA in PAMs. PAMs were transfected with p65-sh1, p65-sh2, p65-sh3 or shN and the cell lysates were subjected to determination of endogenous p65 expression by Western blot at 48 hpt. (e,f) LPS does not inhibit CSFV replication in p65-knockdown PAMs. PAMs treated with LPS were transfected with p65-sh2 and infected with CSFV. CSFV genome RNA and viral titres were determined at 24 and 48 hpi. The data represent the mean ± SD of three independent experiments. **P < 0.01 and ***P < 0.001 calculated using Student’s t-test. ns: not significant.