Fig. 6. FCA autoregulation controls the level of functional transcript γ. (A) Immunodetection of FCA protein. Western blot analysis of total soluble protein extracts from seedlings of Ler and the transgenic line 35S::FCA-gene (a 35S fusion to the FCA gene) in Ler. Filled and unfilled arrowheads denote endogenous and transgenic FCA proteins, respectively. Lower bands are unspecific binding products used as loading controls. (B) Northern blot analysis of poly(A)⁺ RNA isolated from Ler and the transgenic line 35S::FCA-gene in Ler [the same transgene as in (A)]. The blot was probed as indicated in Figure 2C. Numbers denote relative expression and were calculated as in Figure 2C. (C) Quantification of the FCA transcripts levels in the fca-1 mutant. Data are the average of six different experiments and the FCA transcript signals obtained by northern analysis were normalized against that for β-TUBULIN. (D) Northern blot analysis of poly(A)⁺ RNA isolated from Ler, fca-1, and the transgenic lines P_FCA::FCA_{to exon 5}:GUS in Ler and P_FCA::FCA_{to exon 5}:GUS in fca-1. The blot was probed as indicated in Figure 2C. Numbers denote relative expression and were calculated as in Figure 2C.