RNA silencing of RLSB in the C
plant Arabidopsis . Panel A: Morphology of two rlsb-silenced plants. 12-week-old rlsb-silenced Arabidopsis grown on MS media supplemented with 8% sucrose. Panel B: Total soluble proteins isolated from leaves of wild type (Col0) and two representative rlsb-silenced (1 and 2) Arabidopsis plants were separated by SDS-PAGE and silver stained. The positions of LSU and SSU are indicated. Unidentified proteins that were reduced or increased in the silenced plants relative to wild type (open or solid triangles, respectively) are indicated. Panel C: Western blot showing levels of accumulation for LSU and RLSB proteins in leaves from Col0 and two rlsb-silenced Arabidopsis plants. Total maize leaf proteins were separated by SDS-PAGE and analyzed by immunostaining as described in Methods. Panel D: Real-time quantitative PCR showing relative levels of mRNA accumulation for rbcL and RLSB in leaves from Col0 and the two rlsb-silenced Arabidopsis plants. Quantification of transcript levels was standardized to actin mRNA. Data is averaged for four wild type and four mutant siblings, with three repeats run for each of the plant samples. Note differences in scale for panels showing rbcL and RLSB mRNAs. Panel E: Western blots showing accumulation levels of additional proteins in Col0 and the two rlsb-silenced Arabidopsis plants. Protein extracts used for panel C were separated by SDS-PAGE and detected with the indicated antibodies as described. Cp-encoded, chloroplast-encoded proteins (note that CF1 alpha and beta subunits run at the same position on this gel), Nuc-encoded Cp-localized, nuclear-encoded proteins targeted to the chloroplast. Nuc-encoded Mt-localized, nuclear-encoded proteins targeted to the mitochondria. Nuc-encoded Cyto-localized, nuclear-encoded proteins localized within the cytoplasm. For all of the protein and RNA samples analyzed in this Figure, equalization for isolation, loading, and analysis was based on using equalized wet weights of starting leaf material.