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. 2014 Feb 20;8(2):e2707. doi: 10.1371/journal.pntd.0002707

Figure 5. Ufm1 processing activity using fractions derived from Ld1SWT, LdUfsp−/− and add back parasites.

Figure 5

A) Ufm1 processing activity of the fractions of the lysates from Ld1SWT cells obtained after fractionation on Q columns were incubated with the fluorescent substrate containing wild type full length Ufm1 substrate (YFP-Ufm1-ECFP). The reduction in the 530 nm fluorescence from the YFP after 60 min incubation is plotted. Immunoblot showing the distribution of Ufsp protein in the fractions is shown. B). Ufm1 processing activity of the fractions of the lysates from Ld1SWT, LdUfsp−/− and LdUfsp−/−+UfspWT cells obtained after similar fractionation as above were incubated with the fluorescent substrate containing wild type full length Ufm1 substrate (YFP-Ufm1-ECFP). C) Ufm1 processing activity of the fractions of the lysates from Ld1SWT, LdUfsp−/− and LdUfsp−/−+UfspWT cells obtained after similar fractionation as above were incubated with the mutant Ufm1 substrate (YFP-Ufm1G>A-ECFP). Data is representative of three independent experiments. D) Immunoblot of the lysates from Ld1SWT, LdUfsp−/− probed with an anti-Ufm1 antibody.