3 2 pyridyl 5 6 bis 4 phenylsulfonic acid 1 2 4 triazine ferrozine

To study variation in infection-relevant phenotypes between the hybrid isolates, we compared the radial growth of A. nidulans, A. spinulosporus, A. latus, and of all the clinical isolates in different temperatures (30°C, 37°C and 44°C), in the presence of increasing concentrations of oxidative stress-inducing compounds (paraquat and menadione), and on iron starvation. Although the importance of oxidative stress susceptibility is contentious for Aspergillus virulence [102 (link),103 (link)], we chose to examine these phenotypes because it is well established that hosts produce reactive oxygen species in response to infection [104 (link)]. To test for the effects of iron starvation on fungal growth, MM was prepared without any iron source and supplemented or not with 200 μM of the iron chelators Bathophenanthrolinedisulfonic acid (4,7-diphenyl-1,10-phenanthrolinedisulfonic acid [BPS]) (Sigma-Aldrich) and 300 μM of 3-(2-pyridyl)-5,6-bis(4-phenylsulfonic acid)-1,2,4-triazine (ferrozine) (Sigma-Aldrich). For radial growth, isolates were grown in triplicate from 10⁵ spores and incubated at 37°C (except for the temperature test) for 5 days. Growth results in the presence of oxidative stress were expressed as ratio, dividing colony radial diameter (cm) of growth in the stress condition by colony radial diameter in the control (no stress condition).