MOESM4 of Suppression of experimental cerebral malaria by disruption of malate:quinone oxidoreductase

Additional file 4. Deficiency of FH and MQO has no effect on gametocyte production. Blood was obtained from infected mice showing 3% parasitaemia and cultured for 22 h under standardized in vitro culture conditions. Then, mature gametocytes and schizonts were collected by Nycodenz density-gradient centrifugation. (A and B) Expression of gametocyte-specific genes. mdv-1/peg3 [21] and g377 [22] were subjected to semi-quantitative RT–PCR using specific primers (see Additional files 2, 3). The hsp70 was used as a positive control. Samples treated with DNase-treated RNA template (hsp70 (-)) were used as a negative control that is the control of eventual DNA contamination of the RNA preparations. Experiments were performed in duplicate and representative data are shown. (C) Control and Δfh parasites-infected erythrocytes cultured for 22 h. (D) Control and Δmqo parasites-infected erythrocytes cultured for 22 h. White arrows indicate representative mature gametocytes. The scale bars indicate 20 μm. Note that sex-specific features such as nuclear enlargement, the distribution of pigment granules throughout the cytoplasm and enlargement of the cells are observed in both Δfh- and Δmqo-parasite cultures just the same as reported by Mons [23].