Table S1. Gal4 Driver Lines in This Study, Related to s 1, 2, 3, 4, 5, 6, S1, S2, S3, S4, S5, and S6 Genotype Abbreviation Expression patterns Ubi-Gal4 Ubi Ubiquitous dome-gal4;elav-gal80 dome MZ Progenitor-specific in the lymph gland; not expressed in the brain (see S1G,H) ((Ghiglione et al., 2002) Tep4 MZ Progenitor-specific in the lymph gland; not expressed in the brain (see S1I,P) (Avet-Rochex et al., 2010) Antp-Gal4 Antp PSC-specific in the lymph gland (Emerald and Cohen, 2004) Elav-Gal4 Elav Pan-neuronal (Luo et al., 1994) Kurs6-Gal4 Kurs6 Majority of neurosecretory cells in the brain including Kurs6 + GABA + cells; not expressed in the Dilp2 + cells (Siegmund and Korge, 2001) TepIV-Gal4;elavgal80 Hand-Gal4, Hml - HHLT Blood-specific expression including the Gal4 UAS-FLP; lymph gland, circulating hemocytes, and A5C-FRT-STOP- heart (Shim et al., 2012) FRT-Gal4 Hand-Gal4; UAS- HandLT Lymph gland including the PSC and FLP; A5C-FRT- progenitors, and heart (Evans et al., 2009) STOP-FRT-Gal4 Hml-Gal4 Hml Mature blood cell-specific expression in the lymph gland and in circulation (Sinenko and Mathey-Prevot, 2004) Dilp2-Gal4 Dilp2 Insulin-producing neurosecretory cells (Rulifson et al., 2002) Orco-Gal4 Orco Olfactory receptor neuron (ORN) (Larsson et al., 2004) GH146-Gal4 GH146 Projection neuron (PN) (Stocker et al., 1997) GH298-Gal4 GH298 Inhibitory interneuron (iln) (Stocker et al., 1997) OR42a-Gal4 OR42a OR42a (Goldman et al., 2005)
Table S2. Constituents and Concentration of Minimum Odor Media, Related to 6 Nutrient *Sucrose (Fisher Scientific) *Trehalose (Sigma T3663) Glucose solution (Sigma G8644) Amount added per 50ml 0.1 g 0.15 g 0.5 ml 10X Grace s amino acid solution (Sigma G0273) 5 ml (Final 1X) 1 Nitrogen base without amino acids (Sigma Y0626) 0.5 g 2 Cholesterol (Sigma C8667) 2 Phosphatidylcholine (Sigma P3556) 3 Antibiotic-, antimycotic solution (Invitrogen) 10 mg 5 mg Final 1X *Agarose (Invitrogen) 0.5 g (0.1 % w.v) 1 added from 10X stock made in water 2 added from 100X stock made in ethanol 3 added from 100X stock * mix with water and autoclave before adding solutions
Table S3. Quantitation of Phenotypes in s 1, 2, 3, 4, 5, 6, S1, and S2 Genotype Percentages Phenotype 1 F dome MZ -gal4, UAS-2xEYFP 60.7 ± 6.5 Progenitors G dome MZ -gal4, UAS-2xEYFP; UAS- 35.1 ± 7.5 H dome MZ -gal4, UAS-2xEYFP; UAS- 30.9 ± 10.8 RyR RNAi I dome MZ -gal4, UAS-2xEYFP; UAS- 31.5 ± 9.5 Orai RNAi J dome MZ -gal4, UAS-2xEYFP; UAS- 75.5 ± 13 IP3R K dome MZ -gal4, UAS-2xEYFP; UAS- 70.6 ± 15 SERCA DN L dome MZ -gal4, UAS-2xEYFP; UAS- 25.1 ± 12.1 CaMKII RNAi M dome MZ -gal4, UAS-2xEYFP; UAS- 25.1 ± 12.1 CaM B34Q N dome MZ -gal4, UAS-2xEYFP; UAS- 74.8 ± 7.9 CaMKII WT 2 A TepIV MZ -gal4; UAS-GCaMP (Early) 55 ± 12 GCaMP B TepIV MZ -gal4; UAS-GCaMP (Late) 32 ± 6 C TepIV MZ -gal4; UAS-GCaMP; UAS- 34 ± 10 D TepIV MZ -gal4; UAS-GCaMP; UAS- 14 ± 4 E TepIV MZ -gal4; UAS-GCaMP; UAS- 68 ± 9 Serca DN F TepIV MZ -gal4; UAS-GCaMP; UAS- 50 ± 10 Serca DN G TepIV MZ -gal4; UAS-GCaMP; UAS- 16 ± 10 GBR1 RNAi H TepIV MZ -gal4; UAS-GCaMP; UAS- 14 ± 9 GBR1 RNAi I TepIV MZ -gal4; UAS-GCaMP; UAS- 22 ± 7 GBR2 RNAi J TepIV MZ -gal4; UAS-GCaMP; UAS- GBR2 RNAi 9 ± 4 Control dome MZ -gal4, UAS-2xEYFP 60.8 ± 6.5 Progenitors K dome MZ -gal4; UAS-GABA B R1 RNAi 45.7 ± 7.9 L dome MZ -gal4; UAS- 31.4 ± 17.2 M dome MZ -gal4, UAS-2xEYFP; UAS- 61.1 ± 16 GABA B R1 RNAi ; UAS-CaMKII N dome MZ -gal4, UAS-2xEYFP; UAS- GABA B R2 RNA ; UAS-CaMKII 60.2 ± 5.3
3 4 5 6 S1 0.5 ± 0.2 0.53 ± 0.3 75.3 ± 14 84.9 ± 5 Q Antp-gal4, UAS-GFP 1± 0.48 Lymph R Antp-gal4, UAS-GFP; UAS- GABA B R1 RNAi size S Antp-gal4, UAS-GFP; UAS- A elav-gal4 41.3 ± 6 P1+ cells B elav-gal4; UAS-Gad1 RNAi 74.2 ± 5.3 Control Kurs6-gal4, UAS-mCD8GFP 37 ± 15 I Kurs6-gal4, UAS-mCD8GFP; UAS- Gad1 RNAi J Kurs6-gal4, UAS-mCD8GFP; UAS- VGAT RNAi A orco 2 /+ 36 ± 11 Pxn+ cells B orco 2 /Df[orco] 68 ± 16 C Orco-gal4; UAS-hid, UAS-rpr 70 ± 8 D Orco-gal4; UAS-orco RNAi 70 ± 9 E Orco-gal4; UAS- Shi ts1 71 ± 18 Control Kurs6-gal4, UAS-mCD8GFP 32.3 ± 10.3 Pxn+ cells Control GH298-gal4 33.3 ± 10 F GH146-gal4 28.3 ± 8 H GH146-gal4; UAS-Shi ts 74 ± 15 J GH298-gal4; UAS-NaChBac 65.2 ± 8 K Kurs6-gal4, UAS-VGAT; GH146-36.4% ± 11 QF, QUAS-Shi ts Control Kurs6-gal4 32 ± 9.3 Pxn+ cells P Kurs6-gal4; UAS-GABA B R1 RNAi 66 ± 13 O Kurs6-gal4; UAS- 67 ± 16 A Or42a-gal4 26.1 ± 7.5 Pxn+ cells B Or42a-gal4, UAS-Shi ts 67.2 ± 12.9 C Or42a f04305 /Df[Or42a] 70 ± 18 Control GH146-gal normal food 32.3 ± 10.3 Pxn+ cells K GH146-gal4 odorless food 73.2 ± 11.9 P GH146-gal4 odor rescue 37.2 ± 13.5 J HHLT-gal4 57 ± 11 Pxn- cells K HHLT-gal4; UAS- 19 ± 13.3 L HHLT-gal4; UAS-RyR RNAi 24.2 ± 8.7 M HHLT-gal4; UAS-PLC RNAi 26 ± 13 N HHLT-gal4; UAS-CaM B34Q 16.6 ± 11.2 P TepIV MZ -gal4 38.1 ± 7.8 TepIV+ cell Q TepIV MZ -gal4; UAS- 23.8 ± 9 R TepIV MZ -gal4; UAS-RyR RNAi 21.9 ± 10.9 S TepIV MZ -gal4; UAS-IP3R 59.2 ± 6.1 T TepIV MZ -gal4; UAS-SERCA DN 53.8 ± 9 U TepIV MZ -gal4; UAS-CaMKII RNAi 17.7 ± 11.5 V TepIV MZ -gal4; UAS-CaMKII WT 49.2 ± 12.5
S2 S2V S2W 0.5 ± 0.2 0.53 ± 0.3 1 ± 0.25 0.9 ± 0.26 A TepIV MZ -gal4; UAS-GCaMP 39 ± 7.2 GCaMP B TepIV MZ -gal4; UAS-GCaMP; UAS- 16 ± 9 GABA B R1 RNAi C TepIV MZ -gal4; UAS-GCaMP; UAS- 13 ± 5.2 I HandLT-gal4; UAS-GABA B R1 RNAi 11.5 ± 9.8 Pxn- cells O HandLT-gal4; UAS-GABA B R1 RNAi ; 37.8 ± 17.5 UAS-Orai R HandLT-gal4; UAS-GABA B R1 RNAi ; 75.5 ± 11.5 UAS-CaMKII WT J HandLT-gal4; UAS- 18 ± 13.1 P HandLT-gal4; UAS- ; 48.7 ± 13.7 UAS-Orai S HandLT-gal4; UAS- ; 66 ± 14.3 UAS-CaMKII Early Antp-gal4; UAS-GFP 1 ± 0.48 Size instars Antp-gal4; UAS-GFP; UAS- (Fold) GABA B R1 RNAi Antp-gal4; UAS-GFP; UAS- Late Antp-gal4; gal80 ts 1 ± 0.16 instars Antp-gal4; UAS-GABA B R1 RNAi ; gal80 ts Antp-gal4; UAS- ; gal80 ts