Fig. 3: Blocking translation of sfGFP by morpholino oligo enhances TDMD mediated by the CDS trigger.
From: Translation suppresses exogenous target RNA-mediated microRNA decay
a Schematic of morpholino oligonucleotide blocking translation initiation on the sfGFP mRNA transcript. b Western blot analysis of sfGFP protein level in trigger-expressing HEK293T cells with morpholino oligos treatment. HSC70 is used as a loading control. NT: Non-targeting. c RT-qPCR analyses of sfGFP mRNA levels in cells as in b. sfGFP mRNA levels are normalized to GAPDH and the value in cells expressing CDS trigger without morpholino oligos treatment is set to 1. Data are presented as mean ± SD (n = 3 biological replicates). Unpaired two-sided t-tests, ns, not significant. d The influence of morpholino oligos and neddylation inhibitor MLN4924 on miR-16. Representative northern blot analyses of miR-16, miR-7 and miR-92a in cells stably expressing sfGFP reporters with the CDS or 3ʹ UTR trigger. miR-7 serves as a positive control for MLN4924 treatment, which inhibits the ZSWIM8-mediated ubiquitination. miR-92a serves as a loading control. Relative miR-16 levels are shown as mean ± SD (n = 3 biological replicates). e The quantification of northern blots in d. Error bars represent SD, unpaired two-tailed t test calculated the p-values, ns, not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. p(lane 1/lane3)<0.0001 (****), p(lane2/lane3)<0.0001 (****), p(lane3/lane6)<0.0001 (****) and p(lane12/lane9)<0.0001 (****). Source data are provided as a Source Data file.
