Figure 2.

RNase-L interacts with LNX. (A) 293T cells were transfected with pCMV3-RNL-myc, pcDNA3-hLNX-Flag, or the respective empty vectors using Lipofectamine 2000 (Invitrogen). Cell lysates were immunoprecipitated (IP) using anti-Myc-tag or IgG control antibodies bound to protein A/G agarose beads. Bound protein complexes were detected by Western blot analysis using the antibodies as indicated; (B) RNase-L and (C) LNX deletions were used in IP studies as in (A) to determine their respective interaction domains [43,127]. RNase-L deletions were subcloned from the pGEX4T3 plasmid (kindly provided by R.H. Silverman) into the eukaryotic expression plasmid pCMV3B using the BamHI and XhoI restriction sites.