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. Author manuscript; available in PMC: 2012 Sep 8.
Published in final edited form as: Oncogene. 2011 Aug 1;31(10):1207–1216. doi: 10.1038/onc.2011.325

Figure 2.

Figure 2

An inverse correlation between MnSOD activity and percentage of S-phase cells. A. A semi-quantitative RT-PCR assay was used to measure MnSOD and GAPDH mRNA levels in exponential and quiescent cultures of MCF-10A human non-malignant mammary epithelial cells. Primers were designed from the coding sequence to measure total MnSOD mRNA levels (top panel), while the abundance of the two transcripts were assessed by designing primers specific to the 3'-UTR. The primer pairs for the 4.2 kb MnSOD transcript were designed from the sequence between the two PAS sites. The reverse primer for the 1.5 kb MnSOD transcript was designed to anchor the first PAS. PCR-amplified products were separated by agarose gel electrophoresis and visualized by staining with ethidium bromide. B. MnSOD mRNA levels in the same cDNA pools were further analyzed by using a quantitative RT-PCR assay using primers representing the coding sequence. C. Immunoblot analysis of MnSOD in cells representative of different percentage of S-phase. Blots were scanned and quantitated using AlphaImager 2000 and ImageJ software. Fold change calculated first by normalizing to actin levels in individual samples and then relative to MnSOD protein levels in cells with 15 percent S-phase. D. Total cellular proteins were used for biochemical measurements of MnSOD activity.