1
nature research | life sciences reporting summary
June 2017
Corresponding author(s):
Catherine E. Schretter,
Sarkis K. Mazmanian
Initial submission
Revised version
Final submission
Life Sciences Reporting Summary
Nature Research wishes to improve the reproducibility of the work that we publish. This form is intended for publication with a
ll accepted life
science papers and provides structure for consistency and transparency in reporting. Every life science submission will use thi
s form; some list
items might not apply to an individual manuscript, but all fields must be completed for clarity.
For further information on the points included in this form, see
Reporting Life Sciences Research
. For further information on Nature Research
policies, including our
data availability policy
, see
Authors & Referees
and the
Editorial Policy Checklist
.
`
Experimental design
1. Sample size
Describe how sample size was determined.
Sample size was chosen based on preliminary experiments and literature in the
field.
2. Data exclusions
Describe any data exclusions.
No data was excluded.
3. Replication
Describe whether the experimental findings were
reliably reproduced.
All attempts at replication were successful under the experimental conditions
defined.
4. Randomization
Describe how samples/organisms/participants were
allocated into experimental groups.
After eclosion, virgin female flies with the same genotype were sorted into groups
of 10-15 flies per vial at random. All flies in each vial were administered with the
same treatment regime. For each experiment, the experimental and control flies
were collected, treated, and tested at the same time.
5. Blinding
Describe whether the investigators were blinded to
group allocation during data collection and/or analysis.
Blinding was not performed as almost all data acquisition and analysis was
automated.
Note: all studies involving animals and/or human research participants must disclose whether blinding and randomization were us
ed.
6. Statistical parameters
For all figures and tables that use statistical methods, confirm that the following items are present in relevant figure legend
s (or in the
Methods section if additional space is needed).
n/a
Confirmed
The exact sample size (
n
) for each experimental group/condition, given as a discrete number and unit of measurement (animals, litters, cultures, etc.)
A description of how samples were collected, noting whether measurements were taken from distinct samples or whether the same
sample was measured repeatedly
A statement indicating how many times each experiment was replicated
The statistical test(s) used and whether they are one- or two-sided (note: only common tests should be described solely by name
; more
complex techniques should be described in the Methods section)
A description of any assumptions or corrections, such as an adjustment for multiple comparisons
The test results (e.g.
P
values) given as exact values whenever possible and with confidence intervals noted
A clear description of statistics including central tendency (e.g. median, mean) and variation (e.g. standard deviation, interq
uartile range)
Clearly defined error bars
See the web collection on
statistics for biologists
for further resources and guidance.
2
nature research | life sciences reporting summary
June 2017
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Software
Policy information about
availability of computer code
7. Software
Describe the software used to analyze the data in this
study.
Daily activity measurements were taken using Drosophila activity monitors (DAMS,
Trikinetics) and actograms were generated using the ActogramJ software. For
video-assisted tracking experiments, Ethovision or the Caltech FlyTracker (http://
www.vision.caltech.edu/Tools/FlyTracker/) software were used to process videos.
Bout analysis was subsequently performed using simple custom python scripts,
which are available upon request. Gait analysis was performed using the FlyWalker
software package (http://biooptics.markalab.org/FlyWalker/). All statistical analysis
was performed using Prism Software (GraphPad, version 7).
For manuscripts utilizing custom algorithms or software that are central to the paper but not yet described in the published li
terature, software must be made
available to editors and reviewers upon request. We strongly encourage code deposition in a community repository (e.g. GitHub).
Nature Methods
guidance for
providing algorithms and software for publication
provides further information on this topic.
`
Materials and reagents
Policy information about
availability of materials
8. Materials availability
Indicate whether there are restrictions on availability of
unique materials or if these materials are only available
for distribution by a for-profit company.
No unique materials were used.
9. Antibodies
Describe the antibodies used and how they were validated
for use in the system under study (i.e. assay and species).
No antibodies were used.
10. Eukaryotic cell lines
a. State the source of each eukaryotic cell line used.
No eukaryotic cell lines were used.
b. Describe the method of cell line authentication used.
No eukaryotic cell lines were used.
c. Report whether the cell lines were tested for
mycoplasma contamination.
No eukaryotic cell lines were used.
d. If any of the cell lines used are listed in the database
of commonly misidentified cell lines maintained by
ICLAC
, provide a scientific rationale for their use.
No commonly misidentified cell lines were used.
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Animals and human research participants
Policy information about
studies involving animals
; when reporting animal research, follow the
ARRIVE guidelines
11. Description of research animals
Provide details on animals and/or animal-derived
materials used in the study.
For our studies, we used the model organism, Drosophila melanogaster. Majority
of the experiments used 7-day-old adult virgin female flies, except for Extended
Data Figure 1b in which 7-day-old male flies were also used. The fly lines used
include: Canton-S (#64349), Imd-/- (#55711), UAS-dTrpA1 (#26264), Tdc2-GAL4
(#52243), T
β
h-GAL4 (#48332), Th-GAL4 (#8488), Ddc-GAL4 (#7009), Gad1-GAL4
(#51630), ChAT-GAL4 (#60317), Elav-GAL4 (#46655), UAS-T
β
hRNAi (#27667), UAS-
DTI (#25039), and pBDPG4U-GAL4 (#68384) lines from Bloomington Drosophila
Stock Center at Indiana University. Other fly stocks used were OregonR (kindly
provided by A. A. Aravin and K. Fejes T
ό
th), T
β
HM18 (kindly provided by M. H.
Dickinson) Tdc2R054, and tsh-GAL80 (kindly provided by D. J. Anderson). To
minimize the effect of genetic background on behaviors, mutant fly lines were
outcrossed for at least three generations onto a wild-type background.
Policy information about
studies involving human research participants
12. Description of human research participants
Describe the covariate-relevant population
characteristics of the human research participants.
This study did not involve human research participants.