Figure 4.

Ex-circRNAs are associated with translational machinery. Sucrose density gradient analysis of fractions from leaves infected with HSVd (A) or ELVd (B). The position of the 80S monosomes is indicated to separate the polysome fractions from the cytosolic ones. Total RNAs extracted from fractions 2 and 3 (cytosol), and 10 and 11 (polysomes) of HSV- and ELVd-infected plants (C and D, respectively) were analysed by RT-PCR. Detection of EF1α (in cucumber) and Cyclophilin (eggplant) transcripts was used as endogenous control. An in vitro generated transcript of PSTVd (added to the plant extract before being loaded into the sucrose gradient) was used as negative control. HSVd and ELVd RNA and endogenous controls were detected in all analysed fractions. In contrast, exogenous control (PSTVd) expected to be not associated with polysomes, was only present in cytosolic fractions. Northern blot of RNA isolated from polysome fractions derived from leaves infected with HSVd (E) or ELVd (F). Serial dilutions of monomeric transcripts and total RNAs extracted from infected plants were used as hybridization controls. Analyses were performed in duplicate and only representative results are shown