Measurement of ROS production by human hematopoietic progenitor cells expressing N-RasG12D or H-RasG12V. (A) Representative chemiluminescence trace of superoxide production measured using Diogenes. In some replicates (inset), superoxide production was quenched by addition of SOD at 50 minutes. (B) Summary of fold increase of total light emission due to expression of activated Ras over controls during early hematopoietic development. (C) Using Diogenes, superoxide production in H-RasG12V–transduced cells was measured, with either 5 μg/mL SOD or 100 μg/mL catalase present from the start of the assay (n = 3). Bar chart represents total superoxide produced (area under chemiluminescence trace). (D) Using Amplex Red, H2O2 production rate was determined over 4 hours. Data represent mean + 1 SD (n ≥ 3). Statistical significance was calculated by ANOVA using Tukey honestly significant differences, Mann-Whitney test, or Student paired t test. †P < .001; *P < .05.
