Fig. 2.

Polysome profiling demonstrates increased SOD2 mRNA translation efficiency in anchorage-independence. A. Polyribosome profiling was carried out after OVCA433 cells were cultured in attached (A) and anchorage-independent (a–i) conditions (0.5 h) and analyzed following sucrose density gradient centrifugation. Four fractions were collected as indicated, RNA extracted and equal quantities of RNA reverse transcribed to assessed for presence of mRNAs in panel B. B. Representative images of SOD2 and ATF4 RT-PCR from RNA isolated from each polysomal fraction demonstrates an increase in the percentage of mRNA in the heavy polysomal fraction 4 in response to anchorage-independence. C. Quantification of relative SOD2 and ATF4 mRNA levels in each fraction demonstrates increased proportion of SOD2 in fraction 4 following culture in anchorage-independent conditions (n = 3; t-test, *P < 0.05; ***P < 0.001).