Inhibitory effect of depleting specific nutrients on translation.

Wild type L. amazonensis cells were exposed to specific nutrient deprivation conditions [PBS, purines, amino acids [31] and glucose] for 4 h. A cycloheximide (CHX) control for translation inhibition is also shown (A). The effects of depriving purines and AA were measured over longer time periods extending up to 4 days (B). Cells were then incubated with 1 μg/ml puromycin for 20 min, extracted, separated over 12% SDS-PAGE and subjected to western analysis using specific antibodies against puromycin. The bottom lane shows the Ponceau stain of the blot, to verify equal gel loads.