(A) Schematic of the dmpi8WT and dmpi8UP transgenes [10] modified to include a premature stop codon in the
open reading frame of dper, 0.9, or both. The transgenes are
based on a dper cDNA/genomic hybrid and include approximately
825bp 5’ upstream of the 0.9 transcription unit, which is transcribed in
the opposite direction to dper. The 3’ untranslated
regions of both dper and 0.9 overlap by 49bp. Rectangles; black
(dper circadian regulatory sequence), gray (untranslated
regions), green (coding region); black lines (introns). (B-G) Young adult male
(B-E) or female (F-I) flies were exposed to 4 days of 12hr light/12 hr dark
cycles (LD) followed by three days of continuous light (LL) at 25°C. For
each genotype, graphs represent data from 96 individual flies (three independent
transgenic lines × 32 individual flies/line) and data pooled to get the
population average. Shown are the relative sleep levels during the last 2 days
of LD followed by 2 days of LL for transgenic flies carrying the dmpi8UP (red)
or dmpi8WT (blue) versions of the wildtype per-0.9 (B, F) per-STOP (C, G),
0.9-STOP (D, H) or double stop (E, I). Grey rectangle, 12 hr dark period. The
following lines were used to obtain group averages; dmpi8WT, kx-f4c, m38-k, f9;
dmpi8UP, m17, m32, f13; per-STOP[dmpi8WT], m53, f18, f46; per-STOP[dmpi8UP],
m55, m131, m138; 0.9-STOP[dmpi8WT], m50, m62, m73; 0.9-STOP[dmpi8UP], m55, f50,
f59; per/0.9-STOP[dmpi8WT], m94, m100, f38; per/0.9-STOP[dmpi8UP], m42, m81, f5.
Representative examples are shown and similar results were obtained with other
independent lines (see STAR Methods and
KEY RESORCE TABLE for identity of
lines analyzed). See also Figures S1 and S2 for additional results.