Extended Data Fig. 3: Analysis of TaVRN1 transcription in mature pollen grains and stability of TaVRN1 transcripts.
a, Micrograph of a mature pollen grain. Over twenty pollen grains were examined, and a representative micrograph is shown. VN, vegetative nucleus; SN, sperm nuclei. b, TaVRN1 expression in stamens (without pollen grains) and mature pollen gains. Transcripts were quantified by RT-qPCR and normalized directly to TaACT. Data are presented as mean values ± s.d. of three biological replicates. c, Ct (Cycle threshold) values of qPCR. Levels of TaVRN1 and three reference genes including TaACT, TaTUB2 (TUBULIN2) and TaPP2A (PROTEIN PHOSPHATASE SUBUNIT 2A) were quantified in stamen tissues and mature pollen grains. The expression levels of the three reference genes in the haploid pollen grains were significantly lower compared to the stamen tissues. Data are presented as mean values ± s.d. of three biological replicates d, Level of the unspliced nascent transcripts of TaVRN1 is relatively high in mature pollen grains. A promoter region of TaVRN1 serves as a control of genomic DNA. Three biological replicates (Rep) were examined. e-f, Stability analysis of spliced (e) and unspliced (f) TaVRN1 transcripts in late-stage stamens. Cordycepin was used to inhibit transcription. Transcripts were quantified by qPCR and directly normalized to TaACT. Data are presented as mean values ± s.d. of three biological replicates. P values are calculated using two-way ANOVA.
