The expression profiles of E2FA and E2FB are distinct in the developing siliques, but overlap in the proliferation phase. (A) qRT-PCR analyses of the G2- and M-phase-specific CDKB1;1 and the seed maturation LEC2 and WRI1 genes in the developing siliques of the wild-type (WT) at four silique developmental stages (S1-S4, pictured in Fig. S1). (B) The transcript levels of the three E2Fs, namely E2FA, E2FB and E2FC, and the single RBR genes were also analysed in these silique samples by qRT-PCR. Values represent fold-changes normalised to the value of the S1 silique stage (set arbitrarily at 1). Data are mean±s.d., n=3 biological repeats. *P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001 (two-tailed, paired t-test between consecutive silique stages). ns, non-significant. (C) To follow the accumulation levels of RBR, E2FA, E2FB and DPB proteins in developing siliques (S1-S4) specific antibodies were used in immunoblot assays as indicated. The Ponceau-stained proteins were used as loading control. Arrowheads indicate the corresponding E2FA and DPB proteins; arrow marks a slower migrating form of DPB in S4 silique stage. Molecular weights of the specific proteins are shown on the left.