miR-155* targets IRAKM and miR-155 targets TAB2. Human PDCs were transfected with (A,D) mimic control (ctrl), miR-155 mimic (miR-155), miR-155* mimic (miR-155*), IRAKM siRNA (si-IRAKM), or TAB2 siRNA (si-TAB2), (B,E) oligonucleotide control (oligo-ctrl), as-miR-155, or as-miR-155*, and then stimulated with R837 for 16 hours. qRT-PCR analysis of IRAKM and TAB2 mRNA expression was normalized to Rpl13a RNA levels. These graphs show the relative expression of IRAKM mRNA (A-B) and TAB2 mRNA (D-E), normalized to the control level. The data are the means (± SD) of triplicate qRT-PCR, representing at least 3 independent experiments, each based on a different PDC preparation. (C) miR-155 directly suppresses IRAKM mRNA through 3′-UTR interactions. The 3′-UTR of human IRAKM was cloned into the psiCHECK-2 vector, downstream of the Renilla luciferase gene. The construct was then cotransfected into HeLa cells with the indicated amount of the miR-155* mimic (miR-155*) or mimic control (ctrl), and the luciferase activity was quantified. The graph shows the relative luciferase activity normalized to the mimic control values. The data are representative of at least 3 independent experiments. (D) miR-155* inhibits IRAKM protein expression, and miR-155 inhibits TAB2 protein expression. (Top) Western blot analysis of IRAKM protein expression in THP-1 cells 48 hours after transfection with IRAKM siRNA (si-IRAKM), miR-155* mimic (miR-155*), or mimic control (ctrl). (Bottom) Western blot analysis of IRAKM protein expression in THP-1 cells 48 hours after transfection with TAB2 siRNA (si-TAB2), miR-155 mimic (miR-155), or mimic control (ctrl). α-tubulin is shown to confirm equal loading. The data are representative of at least 3 independent experiments, each based on a different PDC preparation.