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. 2013 Feb 25;8(2):e57001. doi: 10.1371/journal.pone.0057001

Figure 4. TbMCM-BP is required for VSG silencing in BF T. brucei.

Figure 4

(A) Diagrams of genes and their chromosomal locations: silent BES-linked VSGs (VSG3 and VSG13), a minichromosomal VSG24, the active VSG2 and BSD marker, and two housekeeping control genes (TUB and URA3). Black circles are telomere repeats. (B) TbMCM-BP deficiency derepresses silent VSG expression. After 24 and 48 hr of post-Cre-induction, mRNA levels of each genes described in (A) were measured and normalized to TbURA3 mRNA level. Fold changes relative to Cre-uninduced cells (0 hr) are plotted. Standard deviations are shown as error bars. (C) Expression of silent VSG13 protein in the VSG2 active BF cells. Cells were fixed with 1% paraformaldehyde and permeabilized with 0.2% NP40. Samples were incubated with chicken anti-VSG2 and rabbit anti-VSG13, and then incubated with secondary antibodies (green for VSG2 and red for VSG13). DNA was stained with DAPI.