Figure 2

Specificity of αT2ib function upon ectopic mouse TLR2 expression. A-D, Assay of NF-κB driven reporter gene activation in HEK293 cells transiently coexpressing mTLR2 (A), mTLR4-MD-2 (B), hTLR3 (C) or mTLR9 (D) and αT2ib (black columns) or αVR-ib (white columns) and challenged as indicated (unstim, non challenged). (E) 3 × 10⁴ human embryonic kidney (HEK) 293 cells per single well of a 6-well microtitre plate transfected with DNA plasmid driving expression of N-terminally-Flag-tagged human TLR2 were cotransfected with graded amounts of TLR2 specific intrabody (αT2ib) or control intrabody (αVR-ib) expression plasmid (line blue, 1 μg; green, 2 μg; red, 5 μg; line orange, 5 μg αT2ib expression plasmid, no primary antiserum; line purple, empty vector) to overexpress each of the two different intrabodies transiently. Upon 48 h cells from each well were subjected to flow cytometrical analysis using Flag-specific primary antiserum and PE coupled secondary antibody.