Fig. 1
Misshapen is a Moesin kinase required for collective cell migration. a Migration indices determined after the depletion of the different Ste20-like kinases. Candidates were depleted using the RNAi lines indicated in the Supplementary Fig. 1, driven by c306-Gal4. This driver is employed for every subsequent experiment, except when specified. See Supplementary Fig. 1a for additional quantifications and details. b Representative images showing the intensity and distribution of pMoe at the onset of migration in control and Msn-depleted clusters. The pMoe channel is displayed individually as inverted greyscale images. c Quantification of the ratio of pMoe mean fluorescence intensity at the cluster periphery, normalized to the signal between nurse cells, after expression of mCherry RNAi (n = 29), msn RNAi#1 (n = 35), msn RNAi#2 (n = 28), Tao RNAi (n = 19), Slik RNAi (n = 27), and Pak3 RNAi (n = 10). n represents the number of independent BC clusters. Non-significant (ns) p > 0,05, *p < 0.05; ***p < 0.001 (one way ANOVA test coupled with Bonferroni correction). Error bars show s.e.m. d Immunoprecipitated wild type and kinase-dead HA-tagged Msn were used in kinase reactions on the CERMAD domain of Moesin. Reactions were analyzed by autoradiography, Western blots, and Coomassie. e Quantification of the distance reached by BCs at stage 10 (0%, 25%, 50%, 75%, or 100% of the total distance) after the expression of a control RNAi or two independent RNAi against msn, together with eGFP, an RNAi-insensitive form of msn or a kinase-dead RNAi-insensitive form of msn, as indicated. n = 143, 25, 50, 93, 42, 50, 35, 31, and 17, respectively, to the histogram order. n represents the number of independent egg chambers analyzed for the quantification. f Representative images showing the localization of Msn and pMoe in BCs. Their co-localization is highlighted by black arrows in separated channels (shown as inverted greyscale images) and yellow arrows in merged images. Co-localization images were obtained by superimposing the black and white negative images of Msn::YFP and pMoe signals. g Representative images showing the localization of Msn in control clusters or after expression of a dominant negative form of Rab11 (Rab11S25N), displayed as inverted greyscale images. h Schematic representation of border cell cluster with labeling of the different cells and cell interfaces. e Quantification of the ratio between the mean Msn::YFP fluorescence signal at the BCs interface within the cluster (BC/BC) or at the periphery of the cluster (BC/NC interface) (n = 15 independent BC cluster, for both condition. ***p < 0.001 (unpaired Student’s t-test). Error bars show s.e.m. i Cropped regions from panel c showing the localization of Msn on vesicles (yellow arrows) in control BCs and Rab11S25N expressing clusters,
