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Figure 1 | Cell Communication and Signaling

Figure 1

From: Structural and functional characteristics of cGMP-dependent methionine oxidation in Arabidopsis thaliana proteins

Figure 1

Protein oxidation assay. OxiSelect™ Intracellular ROS assay kit (Cell Biolabs, Inc. San Diego, CA) was used in the in vivo oxidation experiments according to the assay protocol provided by the manufacturer. Cultured Arabidopsis (Col-0) cells were placed in a black bottom 96-well cell culture plate for 2 h in a shaking incubator. The 2′,7′-dichlorofluorescein diacetate/media solution was added to the cells prior to incubation at 37°C for 1 h. The dye-loaded cells were then treated with 10 μM or 50 μM of cGMP or H2O2. Fluorescence in the cells was measured at 30 and 60 min post-treatment at 480/530 nm using a PHERAstar FS microplate reader (BMG Labtech GmbH, Germany) and the values plotted. Each bar represents data from 3 biological replicates (n = 3), the bars are the standard errors. Treatment with 8-Br-cGMP at the final concentration of 50 μM induces statistically significant differences of the means at p = 0.05 using a two-sample t-test.

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