FIGURE 5: Differentiation in bloodstream form RNAi autophagy mutants. Differentiation of BSF YFP-ATG8.2 ATG5 RNAi mutants (48 h after tet induction) was stimulated by switching cells to differentiation media and incubating at 27oC. Cells were analysed for characteristic PCF markers such as EP-procyclin expression and nuclear/kinetoplast re-configuration.

(A) Western blot of whole cell extracts taken at specific times after induction of differentiation and probed with EP-procyclin, anti-EF1α and anti-OPB.

(B) Fluorescent microscopy of cells 24 h post induction, fixed and labelled with anti-EP procyclin (red) and DAPI (blue) to show changes in the nuclear configuration (nucleus, N and kinetoplast, K). Scale bar 5 µm.

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