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. 2013 Mar 7;9(3):e1003213. doi: 10.1371/journal.ppat.1003213

Figure 3. Analysis of parasites expressing ddFKBPmyc-Rab5A(N158I) and ddFKBPmyc-Rab5C(N153I).

Figure 3

(A) Immunofluorescence analysis of indicated parasites treated for 24 hrs with 1 µM Shld-1 and probed with indicated antibodies (green) and Dapi (blue). (B) Replication assay of indicated parasites grown for 24 hrs in presence, or absence of 1 µM Shld-1 prior to fixation. Average number of parasites per PV was determined. No significant differences in replication were detected. (C) Egress assay of indicated parasites grown for 36 hrs +/− 1 µM Shld-1 before egress was triggered with A23187. Host cell lysis was determined 8 min after induction of egress and normalised with RH hxgprt−parasites. For both mutants the egress is significantly decreased. (D) Invasion assay of indicated parasites treated for 24 h +/− 1 µM Shld-1, scratched and inoculated on fresh HFF cells. Subsequently invasion was determined and normalised with RHhxgprt− parasites. For both mutants the invasion is significantly blocked. (B–D) Mean values and the respective standard deviation of three independent experiments are presented. (***indicates p-value of P≤0.01 and **indicates P≤0.02 in a two tailed Student's test).