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Figure S1. A time series of a living root hair cell from Phaseolus vulgaris responding to NFs from Rhizobium etli. This high-resolution sequence illustrates the transient response in intracellular ROS levels after NFs treatment using non-ratiometric analysis. The transient response commences a few seconds (10–15 sec) after NFs exposure and disappears after 4 min. NFs were added at t = 0 and images were taken every second. Red and blue colors indicate high and blue ROS levels, respectively.

Figure S2. A time series of a living root hair cell from P. vulgaris responding to NFs using a pseudo-ratio-imaging approach. Root hair cells were loaded with both ROS-sensitive and reference dyes for pseudo-ratio-imaging. Cells were then exposed to NFs. The series of images depict transient ROS changes induced by NFs. Note that the intracellular ROS levels at the end of the experiments are higher as compared to images taken before NF addition.

Figure S3. Root hair cells showing the cytoplasmic distribution of the ROS-sensitive (left image) and reference dyes (middle image). The DIC image that depicts the cytoplasm distribution is also included (right image).

Figure S4. Effect of exposure time in root hair cells loaded with a ROS-sensitive dye and a reference dye for ratiometric imaging of intracellular ROS levels.

Exposure times between 10–20 msec allowed constant measurements; however, 50 msec induced a slight increase in control conditions. When exposure times were over 100 msec, further increases were observed (data not shown).

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