Figure 6.

Detection of Ribosome Stalling Is Context Dependent

(A) Reporter construct and expected protein products in the absence or presence of terminal stalling. FLAG-SR-X is a “stalling reporter” in which X represents either a stalling sequence (poly(A), coding for 21 Lysine codons, termed (K^AAA)₂₁) or no insert (termed (K)₀). The RFP:GFP ratio is a quantitative measure of terminal stalling.

(B) Histograms of the RFP:GFP ratios quantified by flow cytometry of WT or ΔZNF598 HEK293 cells expressing the (K^AAA)₂₁ containing reporter. The cells were induced to express the reporter for 22 hr in the presence (red traces) or absence (gray traces) of 10 nM pactamycin, an inhibitor of translation initiation.

(C) HEK293 cells expressing the (K^AAA)₂₁ stalling reporter (left) or control (K)₀ construct (right) were cultured for 22 hr in the absence (shaded gray trace) or presence of varying concentrations of pactamycin (colored traces). Staggered histograms of the RFP:GFP ratio measured by flow cytometry are plotted.

See also Figure S7.