Nature - USA (2020-06-25)

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nature research | reporting summary

October 2018

Field-specific reporting

Please select the one below that is the best fit for your research. If you are not sure, read the appropriate sections before making your selection.

Life sciences Behavioural & social sciences Ecological, evolutionary & environmental sciences

For a reference copy of the document with all sections, see nature.com/documents/nr-reporting-summary-flat.pdf

Life sciences study design

All studies must disclose on these points even when the disclosure is negative.

Sample size No statistical methods were used to pre-determine sample sizes, but our sample sizes were similar to those used in previous publications.

Data exclusions No dataset were excluded from analysis but some exclusion criteria for single neurons were applied when appropriate (e.g. when a neuron

does not have its receptive field where the visual stimulus was presented, it does not make sense to include it in the further steps of the

analysis). A full and detailed section is included in the Method section under 'Inclusion criteria'

Replication Multiple experimental series using both two-photon calcium imaging and electrophysiology were performed. All attempts to replicate the

data were successful.

Randomization Allocation into experimental groups was not randomized. Mice were assigned to a specific experimental group based on their genotype and

all mice types were submitted to a similar visual stimulation protocol and therefore all main findings were reproduced throughout mice

genotypes.

Blinding Data collection and analysis were not preformed blind to the experimental conditions but both collection and analysis were performed by two

different experimentalists.

Reporting for specific materials, systems and methods

We require information from authors about some types of materials, experimental systems and methods used in many studies. Here, indicate whether each material,

system or method listed is relevant to your study. If you are not sure if a list item applies to your research, read the appropriate section before selecting a response.

Materials & experimental systems

n/a Involved in the study

Antibodies

Eukaryotic cell lines

Palaeontology

Animals and other organisms

Human research participants

Clinical data

Methods

n/a Involved in the study

ChIP-seq

Flow cytometry

MRI-based neuroimaging

Animals and other organisms

Policy information about studies involving animals; ARRIVE guidelines recommended for reporting animal research

Laboratory animals Mice of either sex were kept on a C57BL/6 background (except VIP-IRES-cre) and were of the following genotype:

Gad2-IRES-cre (GAD2tm2(cre)Zjh ; JAX:010802) × Ai14 (Gt(ROSA)26Sortm14(CAG-tdTomato)Hze; JAX:007914) for imaging of

layer 2/3 (L2/3) excitatory neurons (9 mice; Fig. 1a-c and 4, Extended Data Fig. 1, 3, and 4); Emx1-IRES-cre (Emx1tm1(cre)Krj ;

JAX:005628) for imaging L2/3 excitatory neurons and axons from LM (5 mice; Fig. 5f-i, and Extended Data Fig. 10a-h); Gad2-IRES-

cre (GAD2tm2(cre)Zjh ; JAX:010802) for imaging L2/3 neurons and labelling inhibitory projections (8 mice; Extended Data Fig. 6,

7h-k, and 10i-k); PV-cre (Pvalbtm1(cre)Arbr; JAX:017320) × Ai14 (Gt(ROSA)26Sortm14(CAG-tdTomato)Hze; JAX:007914) for

imaging of L2/3 parvalbumin-expressing inhibitory neurons (PV; 7 mice; Fig. 2b); VIP-IRES-cre (Viptm1(cre)Zjh; JAX: 010908) ×

Ai14 (Gt(ROSA)26Sortm14(CAG-tdTomato)Hze; JAX:007914) for imaging of L2/3 vasoactive-intestinal-peptide-expressing

inhibitory neurons (VIP; 8 mice; Fig. 2c); Sst-IRES-cre (Ssttm2.1(cre)Zjh; JAX:028864) × Ai14 (Gt(ROSA)26Sortm14(CAG-

tdTomato)Hze; JAX:007914) for imaging of L2/3 somatostatin-expressing inhibitory neurons (SOM; 5 mice; Fig. 2d); Scnn1a-Tg3-

cre (Tg(Scnn1a-cre)3Aibs/J; JAX:009613) and Scnn1a-Tg3-cre (Tg(Scnn1a-cre)3Aibs/J; JAX:009613) × Ai148 (Igs7tm148.1(tetO-

GCaMP6f,CAG-tTA2)Hze; JAX:030328) for imaging layer 4 (L4) excitatory neurons (5 mice and 1 mouse, respectively; Fig. 1d-f,

and Extended Data Fig. 4d); and VGAT-ChR2-EYFP (Tg(Slc32a1-COP4*H134R/EYFP)8Gfng/J; JAX:014548) for electrophysiology

and optogenetic inhibition experiments (20 mice; Fig. 3 and 5a-e, Extended Data Fig. 5, 7a-g, 8, and 9). The mice were housed on

a reverse light cycle (light/dark cycle: 12/12 hrs). At the start of the experiments, all mice were between 2 and 9 months old.

Wild animals This study did not involve wild animals.