Fig. 6: Identification of protein kinases involved in motility.

a Bar chart showing flagellar length: cell body length ratio for 5 protein kinase-deficient mutants identified with significant loss of representation at the top of the transwell, indicating a motility defect. Error bars indicate standard deviation (n = 50 cells for all samples except Δulk4 where there were 43 cells in the analysis). Mutants with a significant difference in flagella:cell body ratio when compared to Cas9T7 were identified using an unpaired t-test with post-Welch’s correction (***Δulk4 p = 1.07e−25; Δstk36 p = 1.93e−27; Δ29.0600 p = 1.01e−22 and Δ02.0570 p = 1.84e−21). b Graph showing the fraction of the population for each promastigote protein kinase mutant progressively swimming. Five sample chambers were prepared for each cell type. The motile fraction of cells in the control (Cas9T7, n = 2532 cells) and four mutant strains: Δulk4 (n = 2936 cells), Δstk36 (n = 4896 cells), Δ29.0600 (n = 2623 cells) and Δ02.0570 (n = 3532 cells). The error bars represent standard error of the mean/95% confidence intervals and the data points denote results from individual chambers. c Schematic showing the protein structures of the four protein kinase mutants identified in the motility screen. d Localisation of the proteins tagged with mNeonGreen. e Bar-seq trajectories for the four protein kinase mutants for Pool 1, EA3 with loss-of-fitness analysed using multiple t-test corrected with post Holm–Sidak method (values are mean ±  S.D., n = 6 biologically independent samples). f normalised bar-seq reads for these four protein kinase mutants during development in the sand fly (values are mean ±  S.E., n = 2 biologically independent samples).