Figure 2.

ZNF598 Is Required to Initiate the RQC Pathway during Poly(A) Translation

(A) (K^AAA)₂₁ cells were transfected with control (blue) or ZNF598 (red) siRNAs for 72 hr, reporter expression induced for 24 hr, and analyzed by flow cytometry (left and middle panels) or immunoblotting for the indicated proteins (right panel). (K)₀ cells (gray) served as a reference.

(B) (K^AAA)21 cells were subjected to the indicated siRNA treatment for 72 hr, reporter expression induced for 24 hr, and analyzed by immunoblotting for the indicated proteins.

(C) (K^AAA)₂₁ reporter expression was induced for 24 hr in wild-type (WT, in blue) and ZNF598 knockout (KO, in red) cells and analyzed by flow cytometry.

(D) (K^AAA)₂₁ cells knocked out for ZNF598 were transiently transfected with the indicated plasmids and the transfected cells (as judged by a co-transfected BFP construct) were analyzed by flow cytometry after reporter induction for 24 hr. The majority of cells displayed restoration of terminal ribosomal stalling to wild-type levels (circled). See also Figure S2.