In vivo evidence of an inverse relationship between SOD1 expression and the level of both S70pBcl-2 and B56δ tyrosine nitration. (A) Western blot (WB) analysis of clinical lymphoma biopsies that are arbitrarily numbered as P1-P14. More complete clinical diagnoses for these samples are described in supplemental Figure 7. Lymphoma biopsies denoted with an asterisk were derived from tumors sufficiently large for further analysis via co-IP assay. (B) Scatter plot analysis of densitometry intensity values of SOD1 and S70pBcl-2. Densitometry values of each sample can be found in supplemental Figure 7. Statistical significance was calculated using Pearson correlation coefficient with a degree of freedom of 18. (C) B56δ was IP from lysates denoted with asterisk and IB for the indicated proteins. (D) This experiment was performed using a rare, huge mantle cell lymphoma that could be sufficiently separated into 3 samples for co-IP analysis. Single cell suspension was prepared, treated for 4 hours with 400 μM DDC, and then harvested for co-IP assay using the indicated Abs. (E) WB analysis of 6 other clinical lymphoma biopsies denoted as P15-P20 with 3 other nonmalignant cell lines: MCF10A breast epithelial cells, HEK293 human embryonic kidney cells, and RWPE-1 prostate epithelial cells. (F) The ratio of S70pBcl-2:SOD1 was calculated using the densitometry value of each protein in supplemental Figure 7 and subsequently correlated with the prognosis or relapse status of each tumor. A ratio greater than 1 would mean a greater abundance of S70pBcl-2 relative to SOD1 and vice versa. Prognosis records, clinical data, and diagnosis of all primary samples can be found in supplemental Figure 7. Three samples—P9, P13, and P20—were diagnosed as reactive lymphoid hyperplasia (not lymphomas) and therefore excluded from the analysis. Clinical records for some patients were unavailable or lost because of a loss of follow-up or some biopsies may have been donated by overseas patients and could not be retrieved.