Figure 1.

Inhibition of myeloid differentiation factor 88 (MyD88) blocks NFκB p65 translocation and hTNFα production. Control shRNA (A) or shMyD88 (B) lentivirally transduced HeLa cells were stimulated with recombinant hTNFα (black bars) or hIL-1β (gray bars) and NFκB localization was determined using anti-p65 conjugated to rhodamine. The bar graphs represent the percent of cells with nuclear p65 localization. The percentage of cells with nuclear p65 was determined in three independent experiments and the SDs between experiments is indicated by the error bars. The number of cells quantified is indicated. (C) THP-1 cells were differentiated and stimulated with the Toll-like receptor (TLR)2 ligand Pam3csK4 (200 pg/mL) overnight. TNFα levels in the supernatants were determined by ELISA. One representative experiment of five independent experiments performed in triplicate, is shown. The two-tailed t-test was used for statistical evaluation.