Figure 8.

Memory experimental design and validation of splicing memory by RT‐PCR. (a) A further characterization of validated AS events was performed in a 45°C assay in order to check AS pattern coherence at a more lethal temperature, test the potential implication of AS in stress memory, and identify high temperature‐specific expression. The 45°C assay was divided in two phases. In Phase I, plants were stressed and sampled on day 1 (T1), day 3( T3), and day 5 (T5) to characterize long responses. At 6 months after the end of Phase I, the plants were subjected to another round of treatment (Phase II) to evaluate the potential acquisition of long‐term splicing memory. (b) Each AS event is represented by a block constituted by two panels. Top panel: transcript model of the small isoform displaying event length, PTC introduction, and where the variation is produced. Bottom panel: RT‐PCR AS patterns in the sampling times corresponding to the 45°C memory assay. The primers used allow the amplification of both splice variants. C, Control; T1, T3, and T5, stressed plant samples after 1, 3, or 5 days of stress.