Figure 5. Detection of protein carbonyl content.

Protein carbonyl groups were labeled with 2,4-dinitrophenylhydrazine and the resulting samples (20 µg of protein) were subjected to western blotting with an antibody against dinitrophenyl: three-day-old cultured epimastigotes (Epi 3d), five-day-old cultured epimastigotes (Epi 5d), five-day-old culturd epimastigotes under nutritional stress (Epi St), adhered epimastigotes after 12 h of differentiation (Adh 12 h) adhered epimastigotes after 24 h of differentiation (Adh 24 h) and metacyclic trypomastigotes (MT). Significant differences between groups are indicated with an asterisk (*). Protein molecular weight is shown on the left. Normalization was performed as described in the Methods. The experiment was carried out three times, with essentially the same results in each case. Results are expressed in densitometric units and are the means of three independent experiments ± SD.