Fig. 4

CSEP0139 and CSEP0182 inhibit BAX-induced cell death in barley. a Suppression of BAX-triggered cell death by CSEP0139 and CSEP0182 in barley. Transient gene overexpression assay was conducted in barley epidermal cells by particle bombardment with combinations of plasmid expressing GFP and empty vector (EV), GFP and BAX and EV, GFP and BAX and CSEP0139, and GFP and BAX and CSEP0182, respectively. Representative pictures indicate that GFP fluorescence shows normal cytoplasmic strands in cells expressing GFP + EV, or GFP + BAX + CSEP0139 or GFP + BAX + CSEP0182, but shows collapsed cytoplasm strands in cell expressing GFP + BAX + EV. Photographs were taken using Nikon A1 confocal laser scanning microscope at 10 h after bombardment. Scale bar = 50 μm. b Quantification of BAX-induced cell death suppressed by CSEP0139 or CSEP0182. The percentage of cells showing collapse of the cytoplasm was shown in various plasmid combinations. The combinations expressing GFP + EV and GFP + BAX + EV were set as controls. The coexpression of GFP and BAX and EV typically induced cell death, thus the percentage of transformed cells showing collapse of the cytoplasm in GFP + BAX + EV was set to 100%. Data shown are means of three replicates ±SE. Means with different letters are significantly different (P < 0.05). Duncan’s multiple range test was used to compare all the means. At least 100 cells were counted per experiment