Fig. 7. Expression of LPG3 mRNA. Northern blot analysis was performed using (A) 5 µg or (B) 15 µg total RNA, and a ³²P-labeled LPG3 coding region probe (0.85 kb EcoRV–BamHI or 1.05 kb NcoI–BamHI fragments, respectively; see Figure 3). The integrity and loading of the RNA is shown by the ribosomal RNA stained with ethidium bromide (bottom). (A) Developmental regulation of LPG3 mRNA. Lane 1, WT promastigotes in logarithmic phase; lane 2, WT promastigotes in stationary phase; lane 3, splenic amastigotes isolated from infected hamsters; lane 4, OB1 promastigotes in logarithmic phase; lane 5, OB1 promastigotes in stationary phase. (B) Effect of stresses on LPG3 mRNA. Total RNA was prepared from logarithmic phase WT promastigotes grown for 24 h at 26°C in either complete medium (lane 1), glucose-free medium (lane 2), complete medium in the presence of 5 µg/ml tunicamycin (lane 3) or for 4 h at 42°C in complete medium (lane 4).