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. 2017 Nov 8;8:1366. doi: 10.1038/s41467-017-01536-x

Fig. 4.

Fig. 4

Mis-localization of Puf118 to the rear of cells inhibits chemotaxis. a Colocalization of GFP-Pla2 and GFP-Lst8 and corresponding mRNAs with myc-tagged TalA-Puf118 (arrowheads) in natural chemotactic streams. Arrow indicates orientation of cell polarity defined by TalA-Puf118 localization. Scale bar, 5 μm. Myc western blot of myc IPs from lysates of cells expressing TalA-Puf118. RT-PCR of indicated mRNAs coimmunoprecipitated with myc-TalA-Puf118. Mouse IgG was used as an immunoprecipitation control. b Chemotactic streams of cells expressing vector control, Puf118-RBD or TalA-Puf118 after starvation for 15 h. Scale bar, 300 μm. c Percent of wild-type cells, cells expressing Puf118-RBD or TalA-Puf118 migrating towards a 0.1 µM/mm (shallow) or 1 µM/mm (steep) cAMP gradient during 180 min. Mean and SD of n ≥ 3 experiments; Mann–Whitney test: *p < 0.05. dg Tracks (d, f) and directionality (e, g) of wild-type cells, cells expressing Puf118-RBD or TalA-Puf118 migrating in a chemotaxis chamber for 180 min in a 0.1 µM/mm (shallow; d, e) or 1 µM/mm (steep; f, g) cAMP gradient. n ≥ 110 cells from ≥3 independent experiments. Mann–Whitney test: ****p < 0.0001, n.s. not significant