Figure 1

Growth and differentiation characteristics of theEATRO1125 trypanosomes used in this study. (A) Growth of the two cultures used for RNA preparation. (B) DNA content during differentiation, measured by FACSScan. The mean and standard deviation are shown, with the number of replicates above the bars. G1 cells had diploid DNA content, G2/M cells 4×, and the S phase intermediate amounts. Cells in the "other" category mostly had more DNA, with peaks at 6× and 8×; a few (under 20% of this category) had less than 2×. (C) Western blots showing expression of various proteins during differentiation. Two blots, A and B, were used; a portion of each, stained for total protein using Ponceau red, is shown at the bottom of the Figure, with molecular weight markers indicated. The arrow points to a thick bloodstream-specific band that migrates at the expected position of VSG. The blot used in each case is indicated on the right. The upper panels show immunoreactivity with: VSG: variant surface glycoprotein Antat1.1; ALD: aldolase; EP: EP procyclin; PAD1: stumpy-specific transporter; RRP6: component of the exosome; TUB: tubulin. (D) Differentiating trypanosomes were stained for VSG and EP procyclin. The percentages of cells with VSG, EP procyclin and both are shown. For the 12 h, 24 h, 48 h and 72 h points, 100 cells were counted. The other samples showed uniform staining over many fields.