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. 2019 Jan 10;176(1-2):306–317.e16. doi: 10.1016/j.cell.2018.10.041

Figure S5.

Figure S5

Oligopeptidase Expression in Trypanosomes, Related to Figure 7

(A) Expression TbPGP or BiPN-TbPGP in vitro. Panels show the growth of parasites ± induction to express BiPN-TbPGP (left panel) or TbPGP (right panel) in vitro (n = 3). In each case expression does not affect the growth of the cells. Error bars = SEM. Inset western blots are shown to confirm protein expression with the BIPN fusion resulting in a larger protein.

(B) Expression of BiPN-TbPOP or TbPOP in vitro. The panels show the growth of parasites ± induction to express BiPN-TbPOP (left panel) or TbPOP (right panel) in vitro (n = 3). In each case expression slows the growth of the cells. Error bars = SEM. Inset western blots show the induced expression of TbPOP and the expresison of the stumpy marker PAD1, which is present on the high density parasites not induced to express TbPOP and the low density parasites induced to express TbPOP.

(C) Western blot demonstrating the extracellular release of TbPOP detected in the culture supernatant (S) with TbPOP in the absence of a BIPN secretory signal; samples were prepared after 1h or 2hr incubation in Creek’s minimal medium without serum (Creek et al., 2013) with TbPOP expression being induced or not with doxycycline. Tubulin remains in the pellet (P) fraction showing that there is no cell lysis.