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. Author manuscript; available in PMC: 2015 Jan 27.
Published in final edited form as: Nat Commun. 2012;3:1312. doi: 10.1038/ncomms2278

Figure 2. PAR-1 serves as a common substrate for FAF and Slimb.

Figure 2

(a–j) Genetic interaction between FAF and Slimb in the retina. All flies were grown at 25°C. Images of female flies are shown. The genotypes are: GMR-Gal4/ +control (a), GMR-Gal4>UAS-Slimb-WT (b), GMR-Gal4>UAS-Slimb-RNAi (c), GMR-Gal4>UAS-Slimb-ΔF (d), GMR-Gal4>UAS-Slimb-ΔF +UAS-PAR-1-RNAi (e), GMR-Gal4>FAFEP381 (f), GMR-Gal4>FAFEP381 +UAS-Slimb-WT (g), GMR-Gal4>FAFEP381 +UAS-Slimb-RNAi (h), GMR-Gal4>FAFEP381 +UAS-Slimb-ΔF (i) and GMR-Gal4>FAFEP381 +UAS-SlimbΔF +UAS-PAR-1-RNAi (j) (n =16, 17, 17, 18, 16,17, 16, 18, 16 and 16 animals, respectively). Statistically significant differences are P<0.001 (GMR-Gal4/ +control, GMR-Gal4>UAS-Slimb-RNAi; GMR-Gal4>FAFEP381, GMR-Gal4>FAFEP381 +UAS-Slimb-RNAi; GMR-Gal4>FAFEP381, GMR-Gal4>FAFEP381 +UAS-Slimb-ΔF; GMR-Gal4>FAFEP381 +UAS-Slimb-ΔF, GMR-Gal4>FAFEP381 +UAS-SlimbΔF +UAS-PAR-1-RNAi ) as determined by Student’s t-test. Experiments were performed in triplicate. Dashed lines outline the eye contour. Values represent areas of retinal surface normalized with GMR-Gal4/ +control. (a–j) Scale bar, 100 μm. (k) Western blot analysis of PAR-1 protein levels in the indicated genotypes. Actin serves as a loading control. Values represent PAR-1 levels in the indicated genotypes normalized with GMR-Gal4>FAFEP381 control in three independent experiments. (l) Double labelling of larval NMJs with anti-horseradish peroxidase (HRP) in green and anti-p-PAR-1 in red. Merged images are shown in lower panels. The genotypes are: Mhc-Gal4>FAF +Slimb-ΔF and Mhc-Gal4/ +control. Scale bar, 5 μm. (m) Quantification of p-PAR-1 signals in l and m after normalization with HRP signal. (n) Quantification of bouton numbers showing genetic interaction between PAR-1 and Slimb at the NMJ. N indicates the number of animals analysed. The error bars represent means±s.e.m. P-values were determined using two-tailed Student’s t-test for each comparison. Experiments were performed in triplicate. (o) Quantification of bouton numbers showing genetic interactions among Slimb, FAF, and PAR-1 at the NMJ. N indicates the number of animals analysed. The error bars represent means±s.e.m. P-values were determined using two-tailed Student’s t-test for each comparison. Experiments were performed in triplicate.

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