Analysis of SBP expression in sbp Arabidopsis mutants in healthy or Cd-stressed conditions. A, SBP transcript level in healthy sbp mutants. RT-PCR was performed on total RNA extracted from 7-d-old wild type and sbp mutants (sbp1, sbp2.5′, sbp3, and the double mutant sbp1sbp3). Results show a representative experiment obtained from two independent sets of data. The number of PCR cycles performed was 27, 27, 35, and 23 for SBP1, SBP2, SBP3, and ACTIN2, respectively. B, Expression analysis of SBP2 in response to Cd in the sbp1 and sbp1sbp3 mutants. RT-PCR was made on total RNA extracted from roots and shoots of 7-d-old seedlings submitted to 0, 50, and 250 μm CdNO3 for 24 h. The number of PCR cycles performed for roots and shoots was 29 and 27, respectively, for SBP1, 31 and 27 for SBP2, and 27 and 23 for ACTIN2. Asterisks show the increased accumulation of SBP2 transcripts in the sbp1 and sbp1sbp3 mutants compared to wild-type seedlings. C, Expression analysis of SBP2 in sbp1 35S∷SBP1 lines L5 and L35 in comparison with wild-type and sbp1 lines transformed with the empty vector pFP101 (lines L6, L8, and lines L31, L33, L34, respectively). RT-PCR analyses were made on total RNA extracted from roots of 7-d-old seedlings. The number of PCR cycles performed was 29 for SBP2 and 25 for ACTIN2. Primers are described in Supplemental Table S1. Asterisks show the increased accumulation of SBP2 transcripts in the sbp1 control lines compared to wild-type control lines.