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. 2021 Jan 27;41(4):689–710. doi: 10.1523/JNEUROSCI.2322-20.2020

Table 1.

Reagents and resources used for this work

Reagent or resource Source Identifier
Antibodies
    Rat polyclonal anti-PDF Depetris-Chauvin et al. (2011) N/A
    Chicken polyclonal anti-GFP Aves Lab Catalog #GFP-1020, RRID:AB_10000240
    Alexa Fluor 647-AffiniPure Donkey Anti-Rat Jackson ImmunoResearch catalog #712-605-150, RRID:AB_2340693
    Cy2-AffiniPure Donkey Anti-Chicken Jackson ImmunoResearch catalog #703-225-155, RRID:AB_2340370
Chemicals
    NaCl, sodium chloride Sigma-Aldrich S7653; CAS: 7647-14-5 (BioXtra, ≥99.5%; AT)
    KCl, potassium chloride Sigma-Aldrich P3911; CAS: 7447-40-7 (ACS reagent, 99.0–100.5%)
    CaCl2.2H2O, calcium chloride dihydrate Sigma-Aldrich 223506; CAS: 10035-04-8 (ACS reagent, ≥99%)
    MgCl2.6H2O, magnesium chloride hexahydrate Sigma-Aldrich M2670; CAS: 7791-18-6 (BioXtra, ≥99.0%)
    NaH2PO4, sodium phosphate monobasic Sigma-Aldrich S8282; CAS: 7558-80-7 (BioXtra, ≥99.0%)
    NaHCO3, sodium bicarbonate Sigma-Aldrich S6297; CAS: 144-55-8 (BioXtra, 99.5–100.5%)
    D-(+)-glucose Sigma-Aldrich G8270; CAS: 50-99-7 (≥99.5%; GC)
    Protease from Streptomyces griseus Sigma-Aldrich P5147; CAS: 9036-06-0 (Type XIV, ≥3.5 units/mg solid, powder)
    Potassium D-gluconate Sigma-Aldrich G4500; CAS: 299-27-4 (≥99%)
    EGTA Sigma-Aldrich E3889; CAS: 67-42-5 (for molecular biology, ≥97.0%)
    HEPES Sigma-Aldrich H3375; CAS: 7365-45-9 (≥99.5%; titration)
    RU-486, mifepristone Sigma-Aldrich M8046, CAS: 84371-65-3 (≥98%)
    Paraformaldehyde Sigma-Aldrich 441244; CAS: 30525-89-4
    NaCl, sodium chloride (for PBS solution) Cicarelli Laboratorios 750; CAS: 7647-14-5
    Na2HPO4, sodium phosphate dibasic Sigma-Aldrich S3264; CAS: 7558-79-4 (for molecular biology, ≥98.5%; titration)
    Triton X-100 Sigma-Aldrich T9284; CAS: 9002-93-1 (BioXtra)
    Goat serum Natocor 734
    Vetbond tissue adhesive 3 M 1469SB
Organisms/strains
    D. melanogaster: w1118 Bloomington Drosophila Stock Center BDSC: 5905; FlyBase: FBal0018186
    D. melanogaster: pdf-GAL: y[1] w[*]; P{w[+mC]=Pdf-GAL4.P2.4}2 Bloomington Drosophila Stock Center BDSC: previously 6900, now available as part of 25031; FlyBase: FBtp0011844
    D. melanogaster: UAS-CD8::GFP: y[1] w[*]; P{w[+mC]=UAS-mCD8::GFP.L}LL5, P{UAS-mCD8::GFP.L}2 Bloomington Drosophila Stock Center BDSC: 5137; FlyBase: FBst0005137
    D. melanogaster: c929-Gal4: w[*]; P{w[+mW. h]=GawB}dimm[929] crc[929] Bloomington Drosophila Stock Center BDSC:25373; FlyBase: FBst0025373
    D. melanogaster: tub-Gal80TS: w[*]; P{w[+mC]=tubP-GAL80[ts]}2/TM2 Bloomington Drosophila Stock Center BDSC: 7017; FlyBase:
FBst0007017
    D. melanogaster: pdfGS: w*; P{UAS-mCD8::GFP.L}LL5; P{Pdf-GS}3/TM3, Sb1 Bloomington Drosophila Stock Center
Depetris-Chauvin et al. (2011) 		BDSC: 80956
FlyBase:
FBst0080956
    D. melanogaster: UAS-dicer2: w[1118]; P{UAS-dicer2, w[+]} VDRC VDRC ID: 60008
    D. melanogaster: R6-Gal4: P{?GawB}crcR6 Helfrich-Forster et al. (2007) FlyBase: FBti0016844
    D. melanogaster: pdf-RFP: Pdf-RFP transgene has 0.6 kb of Pdf regulatory genomic DNA 0.5 kb upstream the start site of transcription and 0.1 kb downstream) fused to DNA encoding mRFP1, a monomeric soluble red fluorescent protein (Shaner et al., 2004). Injected into y w flies. Ruben et al. (2012) FlyBase: FBrf0219602
    D. melanogaster: Ihf01485: PBac{WH}Ihf01485 Exelixis at Harvard Medical School FlyBase:
FBst1017022
    D. melanogaster: Ihf03355: PBac{WH}Ihf03355 Exelixis at Harvard Medical School FlyBase:
FBst1018427
    D. melanogaster: RNAi of cac: P{KK101478}VIE-260B VDRC VDRC ID: 104168
FlyBase:
FBst0476026
    D. melanogaster: RNAi of cac: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02572}attP2 Bloomington Drosophila Stock Center BDSC: 27244; FlyBase: FBst0027244
    D. melanogaster: RNAi of Ca-α1T: P{KK100082}VIE-260B VDRC VDRC ID: 108827
FlyBase:
FBst0480621
    D. melanogaster: RNAi of ClC-a: P{KK101247}VIE-260B VDRC VDRC ID: 110394
FlyBase:
FBst0481966
    D. melanogaster: RNAi of CngA: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02039}attP2 Bloomington Drosophila Stock Center BDSC: 26014; FlyBase: FBst0026014
    D. melanogaster: RNAi of CngA: P{KK108314}VIE-260B VDRC VDRC ID: 101745
FlyBase: FBst0473618
    D. melanogaster: RNAi of Ih: P{KK100190}VIE-260B VDRC VDRC ID: 110274
FlyBase:
FBst0481852
    D. melanogaster: RNAi of Ih: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF03253}attP2 Bloomington Drosophila Stock Center BDSC:29574; FlyBase: FBst0029574
    D. melanogaster: RNAi of Ork1: P{KK107843}VIE-260B VDRC VDRC ID: 104883
FlyBase:
FBst0476711
    D. melanogaster: RNAi of Ork1: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF01926}attP2 Bloomington Drosophila Stock Center BDSC:25855; FlyBase: FBst0025885
    D. melanogaster: RNAi of Shal on the III chromosome NIG Fly Stock Center NIG Stock ID: 9262R-3
    D. melanogaster: RNAi of tipE on the III chromosome NIG Fly Stock Center NIG Stock ID: 1232R-3
    D. melanogaster: RNAi of tipE: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02148}attP2/TM3, Sb[1] Bloomington Drosophila Stock Center BDSC:26249; FlyBase: FBst0026249
    D. melanogaster: RNAi of Atpα: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02910}attP2 Bloomington Drosophila Stock Center BDSC: 28073;
FlyBase: FBst0028073
    D. melanogaster: RNAi of Atpα: P{KK108782}VIE-260B VDRC VDRC ID: 100619
FlyBase:
FBst0472492
    D. melanogaster: RNAi of Calx: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02937}attP2 Bloomington Drosophila Stock Center BDSC: 28306;
FlyBase:
FBst0028306
    D. melanogaster: RNAi of Calx: P{KK109144}VIE-260B VDRC VDRC ID: 104789
FlyBase:
FBst0476622
    D. melanogaster: RNAi of Ca-α1D: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF01848}attP2 Bloomington Drosophila Stock Center BDSC: 25830;
FlyBase:
FBst0025830
    D. melanogaster: RNAi of Ca-α1D: w[1118]; P{GD1737}v51490/CyO VDRC VDRC ID: 51490
FlyBase:
FBst0469449
    D. melanogaster: RNAi of Ca-α1T: w[1118]; P{GD7754}v31963 VDRC VDRC ID: 31963
FlyBase:
FBst0459316
    D. melanogaster: RNAi of Ca-α1T: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02150}attP2 Bloomington Drosophila Stock Center BDSC: 26251;
FlyBase:
FBst0026251
    D. melanogaster: RNAi of eag: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF01471}attP2 Bloomington Drosophila Stock Center BDSC: 31678;
FlyBase:
FBst0031678
    D. melanogaster: RNAi of eag: P{KK107309}VIE-260B VDRC VDRC ID: 100260
FlyBase:
FBst0472134
    D. melanogaster: RNAi of Hk: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02965}attP2/TM3, Sb[1] Bloomington Drosophila Stock Center BDSC: 28330;
FlyBase:
FBst0028330
    D. melanogaster: RNAi of Hk: P{KK109058}VIE-260B VDRC VDRC ID: 101402
FlyBase:
FBst0473275
    D. melanogaster: RNAi of inx2: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02446}attP2 Bloomington Drosophila Stock Center BDSC: 29603;
FlyBase:
FBst0029306
    D. melanogaster: RNAi of inx2: P{KK111067}VIE-260B VDRC VDRC ID: 102194
FlyBase:
FBst0474063
    D. melanogaster: RNAi of Ir: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF01841}attP2 Bloomington Drosophila Stock Center BDSC: 25823;
FlyBase:
FBst0025823
    D. melanogaster: RNAi of Ir: P{KK102249}VIE-260B VDRC VDRC ID: 107389
FlyBase:
FBst0479211
    D. melanogaster: RNAi of Irk2: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF01838}attP2 Bloomington Drosophila Stock Center BDSC: 25820;
FlyBase:
FBst0025820
    D. melanogaster: RNAi of Irk2: w[1118]; P{GD203}v4341 VDRC VDRC ID: 4341
FlyBase:
FBst0465076
    D. melanogaster: RNAi of KCNQ: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02562}attP2 Bloomington Drosophila Stock Center BDSC: 27252;
FlyBase:
FBst0027252
    D. melanogaster: RNAi of KCNQ: P{KK109039}VIE-260B VDRC VDRC ID: 106655
FlyBase:
FBst0478479
    D. melanogaster: RNAi of Ncc69: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF03097}attP2 Bloomington Drosophila Stock Center BDSC: 28682;
FlyBase:
FBst0028682
    D. melanogaster: RNAi of Ncc69: P{KK108763}VIE-260B VDRC VDRC ID: 106499
FlyBase:
FBst0478323
    D. melanogaster: RNAi of Nckx30C: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02574}attP2 Bloomington Drosophila Stock Center BDSC: 27246;
FlyBase:
FBst0027246
    D. melanogaster: RNAi of nrv1: w[1118] P{GD959}v46542 VDRC VDRC ID: 46542
FlyBase:
FBst0466759
    D. melanogaster: RNAi of nrv1: P{KK100406}VIE-260B VDRC VDRC ID: 103702
FlyBase:
FBst0475560
    D. melanogaster: RNAi of nrv2: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF03081}attP2 Bloomington Drosophila Stock Center BDSC: 28666;
FlyBase:
FBst0028666
    D. melanogaster: RNAi of nrv2: w[1118]; P{GD960}v2660 VDRC VDRC ID: 2660
FlyBase:
FBst0456497
    D. melanogaster: RNAi of para: w[1118]; P{GD3392}v6131 VDRC VDRC ID: 6131
FlyBase:
FBst0470199
    D. melanogaster: RNAi of para: P{KK108534}VIE-260B VDRC VDRC ID: 104775
FlyBase:
FBst0476611
    D. melanogaster: RNAi of picot: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF01940}attP2 Bloomington Drosophila Stock Center BDSC: 25920;
FlyBase:
FBst0025920
    D. melanogaster: RNAi of picot: P{KK106848}VIE-260B VDRC VDRC ID: 101082
FlyBase:
FBst0472955
    D. melanogaster: RNAi of ppk: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF03250}attP2 Bloomington Drosophila Stock Center BDSC: 29571;
FlyBase:
FBst0029571
    D. melanogaster: RNAi of ppk: P{KK104185}VIE-260B VDRC VDRC ID: 108683
FlyBase:
FBst0480493
    D. melanogaster: RNAi of ppk12: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02027}attP2 Bloomington Drosophila Stock Center BDSC: 27092;
FlyBase:
FBst0027092
    D. melanogaster: RNAi of ppk12: P{KK101805}VIE-260B VDRC VDRC ID: 105131
FlyBase:
FBst0476959
    D. melanogaster: RNAi of ppk25: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02434}attP2 Bloomington Drosophila Stock Center BDSC: 27088;
FlyBase:
FBst0027088
    D. melanogaster: RNAi of ppk25: P{KK109736}VIE-260B VDRC VDRC ID: 101808
FlyBase:
FBst0473681
    D. melanogaster: RNAi of ppk28: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02153}attP2 Bloomington Drosophila Stock Center BDSC: 31878;
FlyBase:
FBst0031878
    D. melanogaster: RNAi of ppk28: P{KK106316}VIE-260B VDRC VDRC ID: 100946
FlyBase:
FBst0472819
    D. melanogaster: RNAi of sei: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF01474}attP2/TM3, Ser[1] Bloomington Drosophila Stock Center BDSC: 31681;
FlyBase:
FBst0031681
    D. melanogaster: RNAi of sei: P{KK105733}VIE-260B VDRC VDRC ID: 104698
FlyBase:
FBst0476547
    D. melanogaster: RNAi of Sh: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF01473}attP2/TM3, Ser[1] Bloomington Drosophila Stock Center BDSC: 31680;
FlyBase:
FBst0031680
    D. melanogaster: RNAi of Sh: P{KK109112}VIE-260B VDRC VDRC ID: 104474
FlyBase:
FBst0476332
    D. melanogaster: RNAi of Shal: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02154}attP2 Bloomington Drosophila Stock Center BDSC: 31879;
FlyBase:
FBst0031879
    D. melanogaster: RNAi of Shaw: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02982}attP2 Bloomington Drosophila Stock Center BDSC: 28346;
FlyBase:
FBst0028346
    D. melanogaster: RNAi of Shaw: P{KK108371}VIE-260B VDRC VDRC ID: 110589
FlyBase:
FBst0482154
    D. melanogaster: RNAi of SK: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF02571}attP2 Bloomington Drosophila Stock Center BDSC: 27238;
FlyBase:
FBst0027238
    D. melanogaster: RNAi of SK: P{KK107699}VIE-260B VDRC VDRC ID: 103985
FlyBase:
FBst0475843
    D. melanogaster: RNAi of SLO2: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF03426}attP2 Bloomington Drosophila Stock Center BDSC: 32034;
FlyBase:
FBst0032034
    D. melanogaster: RNAi of stj: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF01825}attP2 Bloomington Drosophila Stock Center BDSC: 25807;
FlyBase:
FBst0025807
    D. melanogaster: RNAi of stj: P{KK101267}VIE-260B VDRC VDRC ID: 108569
FlyBase:
FBst0480379
    D. melanogaster: RNAi of Teh2: w[1118]; P{GD3839}v9037 VDRC VDRC ID: 9037
FlyBase:
FBst0471346
    D. melanogaster: RNAi of Teh2: P{KK112449}VIE-260B VDRC VDRC ID: 104951
FlyBase:
FBst0476779
    D. melanogaster: RNAi of Teh4: w[1118]; P{GD3578}v11621/CyO VDRC VDRC ID: 11621
FlyBase:
FBst0450303
    D. melanogaster: RNAi of Teh4: P{KK110985}VIE-260B VDRC VDRC ID: 102161
FlyBase:
FBst0474030
    D. melanogaster: RNAi of trp: y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.JF01441}attP2 Bloomington Drosophila Stock Center BDSC: 31649;
FlyBase:
FBst0031649
    D. melanogaster: RNAi of trp: w[1118]; P{GD372}v1366 VDRC VDRC ID: 1366
FlyBase:
FBst0451102
    D. melanogaster: UAS-pdf on the 2nd Chromosome Renn et al. (1999) N/A
Software
    ImageJ Schneider et al. (2012) https://imagej.nih.gov/ij/
    Infostat Di Rienzo et al. (2018) https://www.infostat.com.ar/
    ClockLab Actimetrics https://www.actimetrics.com/products/clocklab/
    Rethomics Geissmann et al. (2019) https://rethomics.github.io/
    R R Core Team (2014) https://www.r-project.org/
    Micro Manager Edelstein et al. (2014) https://micro-manager.org/wiki/Download_Micro-Manager_Latest_Release
    pClamp 9 Molecular Devices https://moleculardevices.app.box.com/s/d93nukl3chbo206t33cw5fpabsph6wh4
    Clampfit 10 Molecular Devices https://moleculardevices.app.box.com/s/l8h8odzbdikalbje1iwj85x88004f588
    Origin 8 OriginLab https://www.originlab.com/
    GraphPad Prism8 https://www.graphpad.com/