Protein Phosphatase 2A B'α and B'β promote pollen wall construction partially through BZR1-activated CEP1 in Arabidopsis

To identify the target genes of BZR1 involved in PP2A B’α and B’β regulated pollen wall development, a transcriptome analysis was performed by RNA-sequencing using anthers of stages 6–9 from Col-0, b’αβ, bzr1-1D/b’αβ, and bzr1-1D. The up-regulated and down-regulated genes in bzr1-1D are available in the supplemental data of the manuscript, while the up-regulated and down-regulated genes in b’αβ and bzr1-1D/b’αβ are shown here.

Methods
For RNA-Seq, flower buds at stages 6-9 were used to extract RNA and construct libraries with a TruSeqTM RNA Sample Prep Kit (Illumina, San Diego, CA, USA) following the manufacturer’s instructions. The samples were then sequenced with an Illumina Genome Analyzer HiSeq 2000. The resulting sequence reads were mapped to the A. thaliana genome (http://grch37.ensembl.org/Homo_sapiens/Info/Index) using TopHat software (http://tophat.cbcb.umd.edu). Gene expression levels were determined using reads per kilobases per million reads. The identification of differentially expressed genes between samples was performed through Cuffdiff analysis with differentially expressed genes defined as a 1.5-fold change and a false discovery rate (P_FDR) value < 0.05.