Supplementary Information
Methods
Publicly available whole genome microarray expression data was accessed through ModMine
(http://intermine.modencode.org/) was used (Contrino et al., 2012; Graveley et al., 2011). First,
Drosophila orthologs to human PA metabolism genes were identified using Basic Local Alignment Search
Tool (BLAST) (Altschul et al., 1990). Gene expression data for these orthologs was then isolated, meancentered and standard deviation normalized. Polyamine metabolic genes were hierarchically clustered in
Matlab using the Spearman average distance (Howe et al., 2010). Principal component analysis (PCA)
was conducted in Matlab. Metabolite concentrations were extracted from published values measured in
the embryo (Callaerts et al., 1992) and larval/pupa (Dion and Herbst 1970) using an online plot digitizer
(http://arohatgi.info/WebPlotDigitizer/).
The fly strains used were: GAL4-btl, UAS-Act:GFP (Bloomington Stock Center) and E-cad::GFP (Huang
et al., 2009). Antibody stainings of eye imaginal discs and embryos were conducted as previously
described (Halder et al., 1998; Zartman et al., 2009). Polyamines were visualized using an antibody to
spermidine/spermine (Spd/Spm) (Abcam). Tracheal lumen (Gasp) was visualized using 2A12
(Developmental Studies Hybridoma Bank). Isotype controls of mouse and rabbit IgG (Vector Labs)
yielded no signal (data not shown). Secondary antibodies were goat anti-rabbit IgG 561 (Invitrogen), and
goat anti-mouse IgG 647 (Invitrogen).
Imaging was performed on a Nikon Eclipse Ti confocal microscope (Nikon Instruments Inc.) with a
Yokogawa spinning disc (Andor Technology). Image data was collected on an iXonEM+ cooled CCD
camera (Andor Technology) using MetaMorph® v7.7.9 software (Molecular Devices). Shown are
maximum intensity z-projections.
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