2 methyl 1 4 naphthoquinone menadione

To examine the effect of growth under oxidative stress conditions, WT and mutant strains were grown on CM plates containing the oxidants hydrogen peroxide (H₂O₂, 30% in water; Fisher), 2-methyl-1,4-naphthoquinone (menadione; Acros Organics) or diamide [azodicarboxylic acid bis (N,N-dimethylamide) (Sigma)] solutions, at the concentrations shown, following Huang and colleagues [10] (link). Pictures were taken at 5 days post treatment.
To observe the accumulation of H₂O₂ at infection sites, infected rice leaf sheaths (48 hpi) were stained with 3, 3’-diaminobenzidine (DAB, Sigma) as described previously by Chi et al. [11] (link). Rice sheaths were incubated in 1 mg/ml DAB solution in the dark at room temperature for 8 hours and destained with ethanol: acetic acid solution (94:4 v/v) for 1 hr. Samples were excised and observed under a light microscope.
Mutant strains were tested for cell wall and osmotic stress conditions. Cell wall stress condition was performed by adding 100 ug/ml of Congo Red (Sigma) to solid complete media (CM). Osmotic stress conditions were applied using 0.5 M NaCl (Sigma) and 1 M Sorbitol (Sigma) in CM. Pictures were taken at 5 days post inoculation.

CDDO-Me was purchased from Toronto Research Chemicals (North York, ON). CDDO-Im was purchased from Tocris (Bristol, United Kingdom). Bovine serum albumin, Tween-20, flavin adenine dinucleotide, 3,3′-methylene-bis (dicoumarol), D-glucose 6-phosphate sodium salt, glucose-6-phosphate dehydrogenase, thiazolyl blue tetrazolium bromide (MTT), and ConA were purchased from Sigma (St. Louis, MO). 2-Methyl-1,4-naphthoquinone (menadione) and digitonin were purchased from Acros (Antwerp, Belgium), 15Δ-PGJ₂ from Cayman Chemical (Ann Arbor, MI), and CdCl₂ from Thomas Scientific (Swedesboro, NJ). Tris, NADP, and EDTA were purchased from BioRad (Hercules, CA), Roche (Indianapolis, IN), and Thermo Fisher Scientific (Waltham, MA), respectively.