The splicing changes in EJC KD can be rescued by overexpression of siRNA-resistant eIF4A3. (A) Western blot showing the expression level of siRNA-resistant FLAG-eIF4A3 (WT) and a mutant that does not form EJC (D401KE402R, Mut) in the background of control and endogenous eIF4A3 KD condition. GAPDH is shown as a loading control. (B) Immunoprecipitation of FLAG-eIF4A3WT and FLAG-eIF4A3Mut in control and eIF4A3 KD cells. One/15th of input was loaded, and the immunoprecipitated samples were blotted for EJC components. GAPDH is used as a negative control. (C) RT-PCR of alternative splicing patterns in control and eIF4A3 KD cells with eIF4A3WT or Mut. Three representative genes were selected, with one constitutive exon (KPNA1) and two alternative exons (MRPL3, ΔΨ >0; C20orf7, ΔΨ <0). The quantification of triplicate experiments are shown below as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001, one-way ANOVA.