Figure 9.

Constitutive and Inducible Silencing of the FtsH12 Protein.

(A) In Arabidopsis, the FtsH12 gene consists of 19 exons and 18 introns. The sequence and position of the target for artificial microRNA silencing are indicated. Numbers represent nucleotide positions relative to the translational start site +1.

(B) Constitutive silencing of FtsH12 resulted in plants with varying degrees of paleness and delayed growth phenotype in the T1 generation. Plants were grown under a 12-h-light/12-h-dark regime for 6 weeks. Both amiRNA constructs produced plants with comparable phenotypes. The identical wild type (Col) plant was used for both the left and right panels. Bar = 2 cm.

(C) Immunoblot analysis of total protein extracts from the amiRNA FtsH12 lines, using the FtsH12 antibody (upper panel), pdNAD-MDH antibody (lower panel), and actin antibody as a loading control (both panels). FtsH12 and actin, as well as pdNAD-MDH and actin, were detected concurrently on the same membrane using secondary antibodies conjugated to different infrared fluorescence dyes (800CW for FtsH12 and pdNAD-MDH, and 680RD for actin). The migration of molecular mass markers is indicated (left). The gel was loaded on an equal protein (15 µg) basis.