Fig 2. CNOT4 regulates PAF1 but not other PAFc components.

(A, B) Endogenous PAFc protein levels were measured in CNOT4-overexpressing or silenced HEK293 cells via Western blot. The silencing efficiency of CNOT4 was measured by RT-PCR (B, bottom). (C) Cells were transfected with the indicated plasmids, and cell lysates were prepared and immunoprecipitated with Anti-Myc. The interaction between mPAF1 and mLEO1 was determined by Western blotting. (D) Myc-mPAF1 WT or 231–535 was transfected along with sicontrol or siCNOT4 in HEK293 cells. Myc-tagged mPAF1 proteins levels were analyzed by immunoblotting, and CNOT4 depletion was measured by RT-PCR (bottom).The arrows indicate the Myc-mPAF1 of WT or mutant.