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. 2023 May 24;193(1):217–228. doi: 10.1093/plphys/kiad307

Figure 3.

Figure 3.

HyPer7 remains specific to H2O2, in vivo. A and B) Representative ratiometric images at the outset and end of imaging time course for seedlings pretreated with either 1 mM l-ascorbate A) or 1 mM NaCl B). Scale bar indicated in A) applies to all images as they were acquired with identical settings. C) Quantification of stromal HyPer7 oxidation in seedlings pretreated with either 1 mM AsA or 1 mM NaCl (osmotic control) prior to imaging stress ****P <<< 0.0001 (Wilcoxon ranked sum test). White triangle indicates per-plastid mean. White triangle indicates sample mean. Individual points reflect individual segmented plastids, box indicates interquartile range with center line indicating treatment median. Lines extending beyond box indicate 1.5× interquartile range. D) Mean HyPer7 ratio, per plastid, ±1 Se relative to the first frame from abaxial pavement cell sensory plastids over an imaging time course. Seedlings were vacuum infiltrated with a syringe and incubated with either 1 mM AsA or 1 mM NaCl (at pH = 5.7) as osmotic control for 30 min prior to imaging. N > 97 plastids coming from multiple independent plants imaged within 2 h of subjective dawn. Data are from the same experiment presented in C). E) Quantification of stromal HyPer7 oxidation in seedlings pretreated with either 20 µM DCMU or mock (DMSO) prior to imaging stress (N > 200 plastids, **P < 0.01, Wilcoxon ranked sum test). White triangle indicates sample mean. Individual points reflect individual segmented plastids, box indicates interquartile range with center line indicating treatment median. Lines extending beyond box indicate 1.5× interquartile range. F) Mean HyPer7 ratio, per plastid, ±1 Se relative to the first frame from abaxial pavement cell sensory plastids over an imaging time course. Seedlings were pretreated with either 20 µM DCMU or mock (DMSO) and imaged within 2 h of subjective dawn. Data are from the same experiment presented in E). Abbreviations: AsA, L-ascorbate; AU, arbitrary units; DCMU, N-(3,4-dichlorophenyl)-N-dimethylurea.