Supplementary Figure 1: Intestinal hrg-7 is a regulator of heme homeostasis.
From: Inter-organ signalling by HRG-7 promotes systemic haem homeostasis
(a) Top: ClustalW alignment of the hrg-7 5’ flanking region with homologous sequences in C. briggsae and C. remanei. White text with black background indicates a conserved nucleotide in all three species. Red box indicates HERE. Bottom: Alignment of the hrg-1 transcriptional regulatory region with the putative hrg-7 transcriptional regulatory region. Red box indicates HERE. White text with black background indicates conserved nucleotides in the HERE. Blue boxes indicate GATA sites. (b) GFP expression in strain IQ7701 (Phrg−7::GFP) grown in 4 or 20 μM heme. Scale bar = 20 μm. Images are representative of three independent experiments. (c) GFP expression in IQ7701 and IQ7702 (Phrg−7mut::GFP) in 4 μM heme. GFP was only detectable in IQ7701. Images are representative of three independent experiments. (d) Schematic showing that endogenous hrg-7 is knocked down by both hrg-7 dsRNA300bp and hrg-7 dsRNAORF, whereas the transgene hrg-7PR is resistant to knockdown by hrg-7 dsRNA300bp but not to knockdown by hrg-7 dsRNAORF. (e) Immunoblot analysis of HRG-7 and HRG-7PR from IQ6011 and IQ7711 fed dsRNA against control vector, hrg-7300bp, and hrg-7ORF. Membranes were probed with polyclonal anti-HRG-7 antibody and then incubated with HRP-conjugated anti-rabbit secondary antibody. ∗ indicates pro-HRG-7. + indicates mature HRG-7. Unprocessed blots are shown in Figure S6. Data is representative of two independent experiments.
