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imb12027-sup-0001-fig_s1.tif949K

Figure S1. Neuropeptide F (NPF) expression in the adult fly brains. (A, B) NPF expression in control flies and in flies with up-regulated NPF. (C, D) NPF expression in control flies and in flies with down-regulated NPF. Scale bar = 100 μm.

imb12027-sup-0002-fig_s2.tif1429K

Figure S2. Neuropeptide F receptor (NPFR1) expression in the adult fly brains. (A, B) NPFR1 expression in control flies and in flies with up-regulated NPFR1. (C, D) NPFR1 expression in control flies and in flies with down-regulated NPFR1. Scale bar = 100 μm.

imb12027-sup-0003-fig_s3.tif521K

Figure S3. Locomotor activities in flies with down-regulated neuropeptide F/its G protein-coupled receptor (npf/npfr1). Larvae and newly emerged adults were kept at 22 °C, then were transferred to 30 °C for blocking npf/npfr1 activities. The locomotor activities in (A), (B) and (E) are for controls and (C) and (F) for treatments with down-regulated npf/npfr1. (D) and (G) indicate the differences of locomotor activities between controls and treatments from ZT6–7 (the activity valley at mid-day) to ZT10–11 (before lights-off). Descriptions of the histograms, x-axis, scale bars and the statistics are the same as Fig. 1.

imb12027-sup-0004-fig_s4.tif336K

Figure S4. Locomotor activity of flies with up-regulated neuropeptide F (npf) or its G protein-coupled receptor (npfr1) under light : dark cycles. (A, B) Locomotor activity of control flies and (C) flies with up-regulated npf. (D, E) Locomotor activity of control flies and (F) flies with up-regulated npfr1. (G, H) Locomotor activity measured from ZT6–7 (the activity valley at mid-day) to ZT10–11 (before lights-off). Descriptions of the histograms, x-axis, scale bars and the statistics are the same as Fig. 1.

imb12027-sup-0005-fig_s5.tif317K

Figure S5. Effect on locomotor activity of loss of function for neuropeptide F (npf)/its G protein-coupled receptor (npfr1). (A, B) Effect on locomotor activity of loss of npfr1 function driven by C929-Gal4 in flies. Transgenic flies were of the same background as the control. (C, D) Effect on locomotor activity of loss of npf function driven by C929-Gal4 in flies. Transgenic flies were of the same background as the control.

imb12027-sup-0006-fig_s6.tif730K

Figure S6. Locomotor activity of female flies with loss-of-function in neuropeptide F (npf)/its G protein-coupled receptor (npfr1) during light : dark cycles. (A, B) Locomotor activity in the control flies and (C) in transgenic flies with down-regulated npf. (D, E) Locomotor activity in the control flies and (F) in transgenic flies with down-regulated npfr1. (G, H) Activity of evening anticipation before lights-off. A significant difference by one-way anova is indicated in lower-case letters. Error bars indicate the standard error.

imb12027-sup-0007-fig_s7.tif481K

Figure S7. Analysis of the 3′ untranslated region of G protein-coupled receptor of neuropeptide F predicts that it is a conserved candidate target of miR-276b. The green regions highlight the similar sequences.

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