Measuring NPC Activity in Arabidopsis Seedlings

The NPC activity in Arabidopsis seedlings was measured according to Kocourková et al. (2011) (link). Seven-day-old Arabidopsis seedlings (five seedlings for each sample) were transferred from liquid MS solution to 1/8 MS medium containing 10 μM AlCl₃, pH 4 and labeled with 0.66 μg ml^-1 of fluorescent PC (BODIPY-PC, D-3771, Invitrogen, USA). Seedlings were incubated on an orbital shaker at 23°C for 2 h. NPC activity in vitro was measured according to Pejchar et al. (2013) (link) using β-BODIPY-PC (D-3792, Invitrogen, USA). The identification of a BODIPY-DAG corresponding spot was based on a comparison with the BODIPY-DAG standard prepared as described earlier (Pejchar et al., 2010 (link)).