FL RNA translation of MLV-ΔNC. (A) Schematic map of the MLV-ΔNC construct. The black box indicates the deletion of the NC domain of Gag. (B) Representative images of ΔNC translation without (upper panels) and with puromycin treatment (lower panels), acquired on a widefield microscope. Merge and MIP are presented. Scale bars are 10 and 1 µm for zoom insets. Arrows indicate untranslating FL RNA (red), translating FL RNA (yellow), and mature Gag (green). (C) Quantification of the MS2 and ST spots per cell. (D) Proportion of translating FL RNA. For all graphs, the mean ± SEM is shown with n = 30 cells. ns = nonsignificant, ****P ≤ 0.0001 (Mann–Whitney test).