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Fig. 1. dnpc1a gene and mutant phenotypes. (A) Phylogenetic tree of yeast,
worm, fly, mouse and human NPC1 proteins determined using the ClustalW
analysis. According to the BDGP prediction
(www.flybase.org),
dnpc1b encodes a 1223 amino acid protein that lacks any signal
peptide. However, another putative start codon 93 nucleotides before the
BDGP-predicted ATG adds a 31 amino acid peptide that contains a predicted
signal peptide and aligns well with the N termini of NPC1 and NPC1L1. (B) The
gene structure of dnpc1a and deletion regions of two dnpc1a
alleles. (C) Aberrant sterol accumulation in dnpc1a mutants observed
using filipin staining. Left column, wild type; right column, dnpc1a
mutants. Top panels show the Malpighian tubules (inset, magnified view);
bottom panels show midguts. (Left) Filipin staining highlights the lumen of
the Malpighian tubules (top, arrow) and the cell-cell boundaries of midgut
cells (bottom, arrow) in wild-type first-instar larvae. In dnpc1a
mutants (right), in addition to the normal sites of sterol, punctate
accumulations of filipin staining are visible (arrowheads) inside Malpighian
tubule and midgut cells.
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