Figure 4: Tracing distinct degradation pathways employed during NMD by monitoring xrFrags.
From: Interrogating the degradation pathways of unstable mRNAs with XRN1-resistant sequences
(a) Model showing the recruitment of NMD-specific decay-inducing factors to activated UPF1 and their contribution to xrFrag accumulation. (b) Western blot analysis of knockdown efficiencies was performed with the indicated antibodies, tubulin served as loading control. (c–e) Northern blots of RNA samples extracted from stable HeLa cell lines transfected with the indicated siRNAs and expressing the indicated reporter constructs. Endogenous 7SL served as control RNA. Mean values of reporter and xrFrag signal±s.d. (n=3) were quantified and for each knockdown condition the PTC values were normalized to the WT. The ratio of xrFrag to reporter mRNA levels is indicated below the graph.
