Fig 6. Isolation of near death experience (nde) extragenic suppressors of nuclear EDS1-induced autoimmunity.
A. Screening scheme used for suppressor mutant isolation. The severe autoimmune line EDS1-YFPNLS #A3 (NLS#A3) was used for EMS-mutagenesis. Mutants were obtained from screening both M1 plants and M2 pools under restrictive temperature conditions (22°C). B. Macroscopic growth phenotypes of 4-week-old nde1 mutant plants (in the EDS1-YFPNLS #A3 background) and control plants. C. Confocal live cell imaging of representative leaf epidermal cells from 4-week-old plants grown at 28°C shifted to 18°C for 24h. D. PR1 (Y1 axis) and EDS1 (Y2 axis) gene expression in the indicated lines measured by qRT-PCR. Plants were grown at 28°C and shifted to 18°C 24h before RNA extraction. Gene expression was normalized to UBQ10. Standard deviation of ≥ 3 biological replicates is shown. Letters indicate statistically significant differences (ANOVA, Fisher’s LSD Post-hoc test, p < 0,05). E. Immunoblot analysis of total protein extracts from the same plants as used in D.