Fig. 3.

Comparisons of MsHO1 transcripts (A), HO-1 protein levels (B and C), and HO activity (D) in alfalfa seedling roots. Five-day-old seedlings were treated or not with 1, 10, and 100 µM SA (SA1, SA10, and SA100), 20 µM haemin (H), 100 µM ZnPPIX alone, or the combination treatments for 12h. The sample without chemicals was the control (C). The transcript quantification test was carried out after 12h of treatment, normalized against expression of two internal reference genes in each sample (A). HO-1 protein expression was analysed by western blotting (B). The number above the band indicates the relative abundance of the corresponding MsHO1 protein compared with that of the control sample. Coomassie Brilliant Blue-stained gels were present to show that equal amounts of proteins were loaded (C). HO activity was also determined (D). Three independent experiments were performed, and the results showed similar trends.