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. 2010 Apr 8;61(9):2383–2393. doi: 10.1093/jxb/erq072

Fig. 4.

Fig. 4.

Temperature sensitivity of cell death induction by transient expression of AtCNGC11/12, empty vector, S58 (AtCNGC11/12:R557C), and S73 (AtCNGC11/12:E519K) in Nicotiana benthamiana. (a) Induction of cell death in N. benthamiana 24, 48, and 80 h after Agrobacterium infiltration, either shifted from 22 °C to 16 °C at 12 h after Agrobacterium infiltration (lower panels) or not shifted (upper panels). Cell death induction was observed in the leaf area expressing S58, but not empty vector (EV) or S73 after the temperature shift. Cell death induced by AtCNGC11/12 was enhanced by temperature shift. Red circles indicate HR development. (b) Quantitative analysis of cell death in N. benthamiana by electrolyte leakage of leaf discs. S58 expression induced cell death after the temperature shift, but not empty vector (EV) or S73. Samples were taken 80 h after Agrobacterium infiltration. (c) RT-PCR analysis of leaf discs from N. benthamiana leaves expressing AtCNGC11/12, S58 or empty vector (EV). The temperature shift did not significantly affect gene expression of AtCNGC11/12 or AtCNGC11/12:R557C in N. benthamiana leaf discs. actin served as a loading control. Samples were taken 24 h after Agrobacterium- infiltration (12 h after the shift). (d) The expression of AtCNGC11/12:GFP and AtCNGC11/12:R557C:GFP was not altered by the temperature shift. The samples were taken 30 h after Agrobacterium infiltration. The fluorescence of the GFP-fusion proteins was monitored by confocal microscopy.