JCS02832 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Sequence alignments of HRM domains. Aligned are sequences of Drosophila melanogaster Fmi (DmFmi), mosquito Fmi homolog (AgFmi), three mouse homologs (mCelsr1, mCelsr2, mCelsr3), and two human GPCRs, parathyroid hormone/parathyroid hormone-related peptide receptor (hPTH/PTHrPR) and calcitonin receptor (hCTR). Identical amino acid residues among the proteins are shown in black boxes. Asterisks indicate conserved residues. Four cysteine residues, which are required for ligand-dependent signal transduction in GPCRs (Asmann et al., 2000), are indicated by **.

• Supplemental Figure 2 -

  Fig. S2. Quantification of EYFP fluorescence in da neurons. EYFP fluorescence of Fmi::EYFP and ΔN::EYFP was quantified in embryos 20-22 hours old. Confocal images of the da neurons in dorsal clusters of embryos, which expressed either Fmi::EYFP or ΔN::EYFP, were captured by using Zeiss LSM510 with X63 oil immersion objective under conditions of identical parameters with 256 steps (8-bit), laser power (1%), detector gain (700), and pinhole size (96 μm). Individual dorsal clusters were manually outlined to obtain pixel intensity (n=15 for each genotype). The cumulative intensity that belongs to each 30 step was estimated and the mean value ± s.d. was plotted on the graph. We excluded the intensities of the step 0-30 because of background noise. Only the intensity of the step 30-59 indicates a statistically significant difference between the cells expressing Fmi::EYFP and those expressing ΔN::EYFP (Welch’s t-test: P<0.01), but other steps have no statistically significant difference. Therefore, the total expression level of Fmi::EYFP and that of ΔN::EYFP in the da neurons were very similar. Genotypes of embryos examined were IG1-2 Gal109(2)80/IG1-2 Gal109(2)80; UAS-Fmi::EYFP/UAS-Fmi::EYFP (Fmi::EYFP) and IG1-2 Gal109(2)80/IG1-2 Gal109(2)80; UAS-ΔN::EYFP/UAS-ΔN::EYFP (ΔN::EYFP).