Quantitative real time-PCR experiments

SAP11 transcript levels in 35S::SAP11_AYWB and 35S::SAP11_MBSP A. thaliana plants were quantified in mature leaves of three independent, 5-week-old plants. Total RNAs were extracted from 100 mg snap frozen A. thaliana leaves with TRI-reagent (Sigma Aldrich) and cDNA synthesis was performed from 0.5 μg total RNA using the M-MLV-reverse transcriptase (Invitrogen). cDNA was subjected to qRT-PCR using SYBR Green JumpStart Taq ReadyMix (Sigma-Aldrich) in a CFX96 Touch Real-Time PCR Detection System (Biorad) using gene-specific primers for the SAP11-homologs and Actin 2 (AT3G18780) (S9 Table).