Canton s

Manufactured by Bloomington Drosophila Stock Center

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The Canton-S is a wild-type strain of the fruit fly Drosophila melanogaster. It is one of the most widely used genetic model organisms in research laboratories. The Canton-S strain is characterized by its standard morphological features and is often used as a reference strain for various experimental studies.

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54 protocols using canton s

Drosophila Strain Maintenance Protocol

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The following Drosophila strains were used in this study: esg-GFP/CyO and UAS-lacZ lines were kindly provided by Dr. Allan Spradling; esg^ts-Gal4 line was generously gifted from Dr. Benjamin Ohlstein; w¹¹¹⁸ (BDSC #3605), Canton-S (BDSC #64349), UAS-bsk^DN (BDSC #6409) and UAS-EGFR^DN (BDSC #5364) were obtained from the Bloomington Drosophila Stock Center. All flies used in this study were mated females unless otherwise mentioned.
Flies were maintained on standard cornmeal-agar medium (the recipe is: 80 g sucrose, 50 g cornmeal, 20 g glucose, 18.75 g yeast, 5 g agar, 30 mL propionic acid, and 1 L water) at 25°C and 60% humidity under a 12/12 h light/dark cycle. Gene overexpression driven by the esg^ts-Gal4 Drosophila line was repressed at 18°C and activated at 29°C.

Yan L., Zhou J., Yuan L., Ye J., Zhao X., Ren G, & Chen H. (2023). Silibinin alleviates intestinal inflammation via inhibiting JNK signaling in Drosophila. Frontiers in Pharmacology, 14, 1246960.

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Assessing Sleep Deprivation Impact on Drosophila Learning

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Canton-S is obtained from Bloomington Drosophila Stock Center and maintained with cornmeal-sucrose-yeast culture medium under 12h day-night shift at 25°C. For evaluating the impact of disturbance on learning and memory, flies at the age of 5 to 7 day after emergence were used. For assessing the effect of protection of A. julibrissin treatment on learning and memory disturbed by sleep deprivation, flies were treated with A. julibrissin of different concentrations, i.e., 0, 4.5, and 18 mg/ml, for 5 days since emergence and subjected to sleep deprivation at the night of the 5^th day for 12h, and then subjected to T-maze for olfactory associative learning at the 6^th day. For sleep deprivation, Drosophila culture bottles were fixed on the orbital shaker (VWR VX2500) at 850 rpm during ZT 13-24. The culture bottles of the control were placed at the same space.

Chang J.S., Liu H.P., Cheng J., Chen C.J., Hwang S.L., Tseng C.C., Hsu L.F, & Lin W.Y. (2019). Albizia julibrissin Ameliorates Memory Loss Induced by Insomnia in Drosophila. Evidence-based Complementary and Alternative Medicine : eCAM, 2019, 7395962.

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Drosophila Rearing and Genetic Stocks

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All flies were reared on standard cornmeal-based fly food. Unless noted otherwise, during developmental and post-eclosion, flies were raised at 25 °C, ~50% humidity and a 12:12 hr light-dark cycle (1400 ± 200 lx white fluorescent light) in humidity and temperature controlled incubators.
The following stocks were used: Canton-S (BL 64349), tub-Gal80^ts (BL 7019), UAS-CD8:GFP (BL 5130), 20XUAS-IVS-mCD8:GFP (BL 32194), UAS-dMBD-R2-RNAi (BL 30481), and UAS-dMBD2/3-RNAi (BL 35347) were obtained from the Bloomington Stock Center (Bloomington, IN). Drosophila virilis (15010–1051.00) and D. yakuba (14021.0261.38) was received from the Drosophila Species Stock Center (San Diego, CA). Cha-Gal80 was a gift from T. Kitamoto and Jay Hirsh generously provided the Tdc2-Gal4 line.

Gupta T., Morgan H.R., Andrews J.C., Brewer E.R, & Certel S.J. (2017). Methyl-CpG binding domain proteins inhibit interspecies courtship and promote aggression in Drosophila. Scientific Reports, 7, 5420.

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Optogenetic Activation of Taste Neurons

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Flies were housed under a 12 h:12 h light:dark cycle at 25°C and 60–70% humidity on cornmeal, oatmeal, yeast and sucrose food. For all experiments 3-6 day old males were used, which were starved for 10-12 h prior to testing, while supplying water via a wet cloth. Flies were then transferred to the arena using an aspirator and left in the arena for 2-10 hours. The following strains were employed: Gr5a-LexA (gift from Kristin Scott [23 (link)]), LexAop-Chrimson ([21 (link)], w1118; P{13XLexAop2-IVS-CsChrimson.mVenus} attP40, Bloomington 55138), Canton S (from A.v.Philipsborn). The flies expressing Chrimson were fed all-trans retinal (ATR, Sigma Aldrich, CAS Number: 116-31-4) for 2-3 days before the starvation period. ATR food was prepared by mixing normal food with ATR to reach a 400 μM solution and then covered with aluminum foil to avoid degradation. Flies fed on ATR food were kept in the dark under aluminum foil cover.

Seidenbecher S.E., Sanders J.I., von Philipsborn A.C, & Kvitsiani D. (2020). Reward foraging task and model-based analysis reveal how fruit flies learn value of available options. PLoS ONE, 15(10), e0239616.

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Drosophila LRRK2 Mutant Aging Study

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For these experiments adult wild type (WT; Canton-S) and LRRK^WD40 mutant (LRRK^ex1, #34750, from Bloomington Stock Center) Drosophila melanogaster (Dm) males were used. Soon after emergence from pupae, WT or LRRK^WD40 mutant males were separated from females. WT and mutant flies were reared on a standard cornmeal-yeast-agar medium in controlled environmental conditions (24-25°C; 60% relative humidity; light/dark = 12/12 hours). Flies ranging 10–15 days in age were tested according to previous experiments [43 (link)].

De Rose F., Corda V., Solari P., Sacchetti P., Belcari A., Poddighe S., Kasture S., Solla P., Marrosu F, & Liscia A. (2016). Drosophila Mutant Model of Parkinson's Disease Revealed an Unexpected Olfactory Performance: Morphofunctional Evidences. Parkinson's Disease, 2016, 3508073.

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Genetic Modifier Screening in Drosophila

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All stocks used for our modifier screen were obtained from the Bloomington Drosophila Stock Center (Indiana, USA) and are listed in Results. The GMR-GAL4; UAS-cindr^RNAi2.21A/ SM5: TM6b line was generated from UAS-cindr^RNAi2.21A transgene [13 (link)] and the GMR-GAL4 driver line [17 (link)]. In addition, we utilized the following stocks: Canton-S, w¹¹¹⁸, UAS-lacZ and UAS-puc (gifts from R. Cagan), and bsk¹ (Bloomington stock number BL-3088), UAS-bsk (BL-9310), cbl^F165 (BL-9676), nopo^excl42 (BL-57335), nopo^Z1447 (BL-57334), puc^H246 (BL-4390), UAS-slpr^WT-HA (BL-58820), Traf4^EY09771 (BL-17600), UAS-Traf6.S (BL-58991) and Uev1a^DG14805 (BL-20440).

Ketosugbo K.F., Bushnell H.L, & Johnson R.I. (2017). A screen for E3 ubiquitination ligases that genetically interact with the adaptor protein Cindr during Drosophila eye patterning. PLoS ONE, 12(11), e0187571.

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Drosophila Genetic Strains for Research

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w¹¹¹⁸ or genetic background strains were used as control lines in this study. Fly strains used include: Canton-S (Bloomington Drosophila Stock Center, Stock 64349), w¹¹¹⁸, SK^-/- (kindly provided by Dr. Patrick Dolph) [44 (link)], Df(Shaker) (kindly provided by Dr. Kyunghee Koh) [45 (link)], UAS-DNK_v4 [40 (link)], UAS-K_v4/Shal [43 (link)], UAS-K_v1/Shaker (kindly provided by Dr. William Joiner), and UAS-SOD1, UAS-SOD2, UAS-Catalase, UAS-NOX-RNAi and UAS-DUOX-RNAi (all kindly provided by Dr. Matthias Landgraf), UAS-GFP-K_v4/Shal [46 (link)], and elav-GAL4, tub-GAL80^ts, UAS-Dcr² (all obtained from the Bloomington Drosophila Stock Center).

Vallejos M.J., Eadaim A., Hahm E.T, & Tsunoda S. (2021). Age-related changes in Kv4/Shal and Kv1/Shaker expression in Drosophila and a role for reactive oxygen species. PLoS ONE, 16(12), e0261087.

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Drosophila RNAi and Overexpression Assays

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Canton S (Bloomington #1), Sgg RNAi (Bloomington #35364), Spalt-LacZ (gift from S. Newfeld), Actin-Gal4, Apterous-Gal4, HH-Gal4, MTD-Gal4 (Bloomington #31777), UAS-MWT-flag, UAS-MadA204/08-flag. All Drosophila crosses were carried out at room temperature 25°C.

Aleman A., Rios M., Juarez M., Lee D., Chen A, & Eivers E. (2014). Mad linker phosphorylations control the intensity and range of the BMP-activity gradient in developing Drosophila tissues. Scientific Reports, 4, 6927.

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Rearing Drosophila Flies in Standard Media

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Flies were reared at 22°C–25°C in standard cornmeal-dextrose media. Wild type flies used in the experiments were Canton-S (BL 64349) obtained from Bloomington stock center. Orco-GAL4 (BL 23292) and UAS-GCaMP6m (BL 42748) flies were also obtained from Bloomington stock center. Following is the composition of the media per 100 food vials: dextrose-100 gms, inactive dry yeast-50 gms cornmeal-70 gms, Drosophila agar-6 gms, propionic acid-6 mL, tegosept-12 mL, water-1025 mL.

Clark J.T., Ganguly A., Ejercito J., Luy M., Dahanukar A, & Ray A. (2022). Chemosensory detection of aversive concentrations of ammonia and basic volatile amines in insects. iScience, 26(1), 105777.

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Genetic Tools for Drosophila Immunity

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dvps35¹ and UAS-dvps35RNAi were described previously (Belenkaya et al., 2008 (link)). w¹¹¹⁸, Canton-S, tubGal4, cgGal4, Tl^r3, Spz², Spz^rm7 (also known as Spz⁴), UAS-SpzRNAi-1 (BL28538), UAS-SpzRNAi-2 (BL34699) and UAS-statRNAi (BL33637) were from Bloomington Stock Center. Tl^RxA was obtained from Dr. K.V. Anderson. YP1-Gal4 was described previously (Hu et al., 2004 (link)). UAS-dvps26RNAi (GD18396), UAS-SPERNAi-1 (KK104906) and UAS-SPERNAi-2 (GD30971) were from VDRC (Dietzl et al., 2007 (link)). For mosaic clone analysis, y w hsp70-flp;FRT^G13 ubiquitin-GFP/FRT^G13 dvps35¹ flies were generated and heat-shocked at 37°C for 2 hours at the beginning of first instar stage. For flip-out clone analysis, y w hsp70-flp/+;AyGal4 UAS-GFP/UAS-Toll^10bMyc flies were generated and heat-shocked at 37°C for 30 minutes at the beginning of first instar stage.

Zhou B., Yun E.Y., Ray L., You J., Ip Y.T, & Lin X. (2014). Retromer Promotes Immune Quiescence by Suppressing Spätzle-Toll pathway in Drosophila. Journal of cellular physiology, 229(4), 512-520.

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