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Genetic dissection of a Leishmania flagellar proteome demonstrates requirement for directional motility in sand fly infections

Fig 1

Isolation of flagella and deflagellated cell bodies.

(A) Overview of the deflagellation and differential centrifugation protocol. Percentage sucrose concentration (w/v) is indicated. (B) Micrographs show merged phase and fluorescence channel (SMP-1::GFP, green; Hoechst DNA stain, red) for each isolation stage (i-iv) depicted in (A). (i) L. mexicana SMP1::GFP cells before deflagellation, (ii) cells after deflagellation, (iii) isolated flagella (F) and (iv) deflagellated cell bodies (CB). Scale bars represent 20 μm. (C) Protein gel stained with SYPRO RUBY. Numbers on the left indicate molecular weight in kDa, numbers below indicate cell equivalent of protein loaded on each lane. Each sample lane of FS, FI, CS and CI was cut into eight pieces and analysed by mass spectrometry (two biological replicates). (D) Venn diagram shows total number of all detected proteins (≥ 2 peptides detected, p-value > 0.95) of both replicates.

Fig 1