(A,B) Percentage of cells in individual brains exhibiting polyploidy in w1118 (A) and Canton-S (B) male and female whole brains. Age in days indicates days post-eclosion. Box plots showing range, …
Source Data for Figure 1A.
Source Data for Figure 1B.
(A–C) Flow Cytometry gating strategy to ensure doublet discrimination. (D) The small population of mz19-GAL4, UAS nGFP labeled neurons are detectable by flow cytometry assay. (E) Dot plots showing …
(A) 14d Canton-S (CS) males reared in 12 h L/D cycles do not show significantly different polyploidy compared to age-matched CS males reared in 24 hr darkness from eclosion. Error bars = mean ± SEM; …
Representative flow cytometry dot plots showing polyploid neuronal (A) and glial (B) cells in 2 week old male brain (A) Neuronal nuclei are labeled using nsyb-GAL4, UAS-nGFP, neuronal cells are …
Source Data for Figure 2C.
Source Data for Figure 2D.
Source Data for Figure 2E.
(A,B) Micrographs showing expression pattern of moody-GAL4 (A) and (Breathless) btl-GAL4 (B) in the adult brain to label SPGs and tracheal cells respectively. (C) Flow cytometry based quantification …
(A) Schematic of ‘early labeling’ using CoinFLP to identify potential cell fusion events. Early CoinFLP labeling will label diploid cells either with GFP or RFP. Any double-labeled cells in an …
(A) Schematic showing labeling protocol for inducing a ‘late-FLP’ in brains where polyploidy is expected to identify polyploid cells in situ. A proportion of cells with multiple copies of the genome …
Source Data for Figure 4C.
Heat-shock induced CoinFLP was used to stochastically label cells in the adult brain with UAS-GFPNLS (magenta), LexAop-RFPNLS (green), or both 24 hr prior to dissection. Optic lobes at 14 days (A–G) …
(A) DAPI integrated intensity was quantified for 132 nuclei from CoinFLP- labeled 14 day optic lobes using FIJI. Nuclear boundaries were guided by lamin staining for Dapi quantifications from at …
Source Data for Figure 4—figure supplement 2 panel A.
Padj = adjusted P value. Upregulated GO terms shown in solid purple (A), downregulated GO terms shown in grey bars outlined with purple (B’). Representative images showing pH2AV foci in 1 (C,C’) and …
(A) Most significantly upregulated GO terms in 7d OL compared to 2d OL. Most significantly upregulated (B) and downregulated (C) GO terms in 21d CB compared to 2d CB. Most significantly upregulated …
Aggregated data based on (A) age (2d vs 21d, n = 6 replicates), sex (B, n = 9 replicates) or brain region (C n = 9 replicates) was compared to previously published datasets. Bar graphs show Mean log …
The list of comparisons and references for validation of RNAseq dataset.
(A) Polyploidy in neurons is not p53 dependent. Percentage of polyploidy under each condition was measured in individual 7d male brains (n = 5). (B) w1118 males treated with 2 mM paraquat (PQ) from …
Source Data for Figure 6D.
Source Data for Figure 6F.
Source Data for Figure 6H.
Source Data for Figure 6I.
Source Data for Figure 6J.
(A) One day and 7d Canton-S brains stained with both Sytox-Green (green) and Propidium Iodide (red) show similar labeling with both cell death markers. (B) Percentage of cells incorporating …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (D. melanogaster) | w1118 | Bloomington Drosophila Stock Center | BDSC 5905 | isogenic |
Genetic reagent (D. melanogaster) | Canton-S | O. Shafer lab | n/a | WT |
Genetic reagent (D. melanogaster) | Oregon-R | C. Collins lab | n/a | WT |
Genetic reagent (D. americana) | Drosophila americana | P. Wittkopp lab | n/a | Non melanogaster Drosophila |
Genetic reagent (D. mauritiana) | Drosophila mauritiana | P. Wittkopp lab | n/a | Non melanogaster Drosophila |
Genetic reagent (D. melanogaster) | glass60J | O. Shafer lab | n/a | Mutant for glass |
Genetic reagent (D. melanogaster) | w;nSyb-GAL4/Cyo | M. Dus lab | n/a | pan-neuronal |
Genetic reagent (D. melanogaster) | w;+;nSyb-GAL4 | M. Dus lab | n/a | pan-neuronal |
Genetic reagent (D. melanogaster) | w;UAS-nGFP;Repo-GAL4, tubulin GAL80TS | Buttitta lab stocks | n/a | pan-glial |
Genetic reagent (D. melanogaster) | w;UAS-nGFP | Buttitta lab stocks | n/a | UAS nuclear GFP |
Genetic reagent (D. melanogaster) | w;+;UAS-nGFP | Buttitta lab stocks | n/a | UAS nuclear GFP |
Genetic reagent (D. melanogaster) | w;Moody-GAL4 | C. Collins lab via Klambt Lab | n/a | Sub-perineurial glia |
Genetic reagent (D. melanogaster) | y,w;mz19-mCD8::GFP | Bloomington Drosophila Stock Center | BDSC 23300 | Antennal lobe projection neuron |
Genetic reagent (D. melanogaster) | w1118;ELAV-GAL4,UAS-nGFP | Bloomington Drosophila Stock Center | BDSC 49226 | pan-neuronal |
Genetic reagent (D. melanogaster) | y,w;breathless-GAL4 | DGRC Kyoto | 105276 | Trachea |
Genetic reagent (D. melanogaster) | w-;GAD1-GAL4/SM6 | O. Shafer lab | n/a | GABAergic |
Genetic reagent (D. melanogaster) | w-;OK371-GAL4,UASn-GFP | Buttitta lab stocks | n/a | Glutamatergic |
Genetic reagent (D. melanogaster) | w;ChaT-GAL4 | O. Shafer lab | n/a | Cholinergic |
Genetic reagent (D. melanogaster) | w1118;+;GMR-12C11-GAL4 | Bloomington Drosophila Stock Center | BDSC 76324 | Tm3a |
Genetic reagent (D. melanogaster) | w1118;+;GMR- 42H01-GAL4 | Bloomington Drosophila Stock Center | BDSC 48150 | Dm9 |
Genetic reagent (D. melanogaster) | w1118;+;GMR-23G11-GAL4 | Bloomington Drosophila Stock Center | BDSC 49043 | Dm4 |
Genetic reagent (D. melanogaster) | w1118;+;GMR-30B06-GAL4 | Bloomington Drosophila Stock Center | BDSC 47529 | Dm10 |
Genetic reagent (D. melanogaster) | w1118;+;GMR-26H07-GAL4 | Bloomington Drosophila Stock Center | BDSC 49204 | Dm2 |
Genetic reagent (D. melanogaster) | y;w;NP3233-GAL4/Cyo | DGRC Kyoto | 113173 | Astrocyte-like |
Genetic reagent (D. melanogaster) | y;w;NP2222-GAL4/Cyo | DGRC Kyoto | 112830 | Cortex glia |
Genetic reagent (D. melanogaster) | w;mz97-GAL4 | C. Collins lab via Klambt Lab | n/a | Wrapping glia |
Genetic reagent (D. melanogaster) | y,w,UAS-mCD8::RFP,LexAop2-mCD8::GFP; CoinFLP-LexA::GAD.GAL4 | Bloomington Drosophila Stock Center | BDSC 59270 and 59271 | CoinFLP |
Genetic reagent (D. melanogaster) | y,w,hs-FLP;LexAop-RFPnls; UAS-GFPnls | Buttitta lab stocks | n/a | hs-FLP used with with CoinFLP nuclear GFP and RFP |
Genetic reagent (D. melanogaster) | ey-FLP | Bloomington Drosophila Stock Center | BDSC 5576 | ey-FLP |
Genetic reagent (D. melanogaster) | y,sev,w;UAS-cdc6RNAi | Bloomington Drosophila Stock Center | BDSC 55734 | cdc6KD |
Genetic reagent (D. melanogaster) | w; UAS-gemininRNAi | Bloomington Drosophila Stock Center | BDSC 30929 and 50720 | gemininKD |
Genetic reagent (D. melanogaster) | w1118;GUS-p53 | Bloomington Drosophila Stock Center | BDSC 6584 | UAS-p53WT |
Genetic reagent (D. melanogaster) | y,w1118; UAS-p53 259N | Bloomington Drosophila Stock Center | BDSC 6582 | UAS-p53DN |
Antibody | anti-ELAV (rat monoclonal) | Developmental Studies Hybridoma Bank | Rat-ELAV-7E8A10 | 1: 100 |
Antibody | anti-pH2AV (mouse monoclonal) | Developmental Studies Hybridoma Bank | UNC93-5.2.1 | 1: 100 |
Antibody | anti-Repo (mouse monoclonal) | Developmental Studies Hybridoma Bank | 8D12 | 1: 100 |
Antibody | anti-Lamin (mouse monoclonal) | Developmental Studies Hybridoma Bank | ADL67.10 | 1: 100 |
Antibody | Alexa Fluor 568 anti-mouse (goat polyclonal) | ThermoFisher | A11031 | 1: 1000 |
Antibody | Alexa Fluor 568 anti-rat (goat polyclonal) | ThermoFisher | A11077 | 1: 1000 |
Antibody | Alexa Fluor 488 anti-mouse (goat polyclonal) | ThermoFisher | A11029 | 1: 1000 |
Antibody | Alexa Fluor 488 anti-rat (donkey polyclonal) | ThermoFisher | A21208 | 1: 1000 |
Other | DAPI | Sigma-Aldrich | D9542 | 1: 1000 |
Other | Dye-cycle violet | ThermoFisher | V35003 | 2: 1000 |
Other | Sytox Green | ThermoFisher | S7020 | 2: 1000 |
Other | Propidium Iodide | Sigma-Aldrich | P4170 | 2.25: 1000 |
A list of antibodies and stains used.
A list of Drosophila stocks used.
Proportions of cell types polyploid in the whole brain.
Proportions of polyploid cell types in the OL.