Fig. 4.

BmMD-2A and BmMD-2B assist BmToll9 to activate LPS-induced AMP gene promoter activity in insect cell lines. Mean relative luciferase activity ± SEM in extracts prepared from cotransfected BmN (A) or S2 cells (B). BmN and S2 cells were cotransfected with pIEx-4–BmToll9–V5 or pIEx-4–GFP–V5 plus pGL3B-moricin, pGL3B-cecropin B, or pGL3B-attacin 1. Dual-luciferase assays were then conducted using extracts prepared from cells in which GFP was expressed with either no factors or LPS added for 12 h (gray bars); BmToll9 was expressed and either no factors or LPS, BmMD-2A–His, or BmMD-2B–His was added for 12 h (blue bars); or BmToll9 was expressed and LPS plus BmMD-2A–His or BmMD-2B–His were added for 12 h (red bars). Three biological replicates were generated per treatment. For each graph, different letters above bars indicate treatments significantly differed from one another (P < 0.05; ANOVA followed by a post hoc Tukey’s honestly significant difference [HSD] test).