Fig. 4: Translation inhibition facilitates TDMD when triggers locate in the coding region.

a Illustration of the molecular mechanisms of six translation inhibitors. Northern blot analyses of miR-16 in ZSWIM8 KO and Scr cells containing the sfGFP CDS or 3ʹ UTR triggers treated with LTM (b) or TG (e) for 8 hours. miR-7 serves as a positive control for ZSWIM8 KO and miR-92a is used as a loading control. Relative miR-16 levels are shown as mean ± SD (n = 3 biological replicates). The influence of triggers by LTM (d) or TG (g) treatment on miR-16 length. Plotted are intensity values as a function of gel migration measured by line densitometry of northern blot in (b) or (e). Peaks correspond to lengths of miR-16 isoforms, as indicated (arrowheads). c and f The quantification of northern blots in b and e, respectively. Unpaired two-tailed t test calculated the p values, ns, not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. In c, p(lane2/lane1) = 0.0053 (**), p(lane4/lane2) = 0.0179(*), p(lane6/lane2) = 0.0123(*), p(lane10/lane8) = 0.0075(**), p(lane12/lane8) = 0.0052(**). In (f) p(lane10/lane8) = 0.0003(***), p(lane12/lane8) = 0.0004(***). h Representative immunoblots detecting puromycin-labeled nascent proteins and sfGFP in HEK293T cells expressing the 3ʹ UTR trigger, treated with six translation inhibitors. Ponceau staining of the blot is used to quantify the total protein in each sample. Source data are provided as a Source Data file.