uterus with two 9-mm electrode paddles positioned on either side of
the head (BTX, ECM830).
Patch-qRT–PCR of NBPF1–GFP plasmid overexpressed cells
Coronal slices containing cells overexpressing the NBPF1–GFP plasmid
were prepared using a vibratome (VT1200S, Leica, Wetzlar, Germany) in
oxygenated (95% O 2 and 5% CO 2 ) ice-cold sucrose-based artificial cerebro-
spinal fluid (s-ACSF, 234 mM sucrose, 2.5 mM KCl, 26 mM NaHCO 3 , 1.25 mM
NaH 2 PO 4 , 11 mM d-glucose, 0.5 mM CaCl 2 and 10 mM MgSO 4 ). The
slices were kept in an incubating chamber filled with oxygenated ACSF
(126 mM NaCl, 3 mM KCl, 1.2 mM NaH 2 PO 4 , 2.4 mM CaCl 2 , 1.3 mM MgSO 4 ,
26 mM NaHCO 3 , 10 mM d-glucose) at 34 °C for 30 min. After a recovery
period of at least 60 min at room temperature, an individual slice was
transferred to a recording chamber and was continuously superfused
with oxygenated ACSF (4 ml/min) at room temperature. We captured
whole cells overexpressing the NBPF1–GFP plasmid and distributed
each into a single tube, and then we used SMART-seq2 to amplify the
mRNA into a cDNA library. Then, we used qRT–PCR to detect NBPF1 and
LHX2 gene expression. Specific genes were amplified using the following
primers: GAPDH forward GTC AAG CTC ATT TCC TGG TAT GAC, reverse
TAT GGG GGT CTG GGA TGG AA; NBPF1 forward GCG AGG CTG CCC GAG
CTT CT, reverse GAC TTC GCG TAA CTT CCC ATT CA; LHX2 forward GAA
CGA TGC TGA ACA CCT GG, reverse AAC CAG ACC TGG AGG AC TCT C.
Statistical analysis
Comparisons between two groups were made using t-tests. The quan-
tification graphs were analysed by using GraphPad Prism (GraphPad
Software). Sample size and P values are given in the Figure legends.
Reporting summary
Further information on research design is available in the Nature
Research Reporting Summary linked to this paper.
Data availability
The scRNA-seq data and ATAC-seq data used in this study have been
deposited in the Gene Expression Omnibus (GEO) under accession
number GSE131258. Raw image files used in the figures that support
the findings of this study are available from the corresponding authors
upon reasonable request.
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Acknowledgements This work was supported by the National Key R&D Program of
China (2019YFA0110100), the Strategic Priority Research Program of the Chinese Academy
of Sciences (XDA16020601, XDB32010100), the National Basic Research Program of China
(2017YFA0102601, 2017YFA0103303), the National Natural Science Foundation of China
(NSFC) (91732301, 31671072, 31771140, 81891001), the Grants of Shanghai Brain-Intelligence
Project from STCSM (16JC1420500), the Grants of Beijing Brain Initiative of Beijing
Municipal Science & Technology Commission (Z181100001518004).Author contributions Q.W. and X.W. conceived the project, designed the experiments and
wrote the manuscript. S. Zhong and Y.L. performed the scRNA-seq experiment. W.D.
performed the ATAC-seq and animal surgery. S. Zhong, Y.L., X.F., S. Zhang and F.T. analysed
the RNA-seq data. S. Zhong and H.D. analysed the ATAC-seq data. L.S. and R.C. collected
single cells by patch-clamping. Z.L. and S. Zhong carried out qRT–PCR. Q.M. prepared the
samples. S. Zhong, Q.W. and W.D. performed immunostaining, in situ hybridization and
imaging. All authors edited and proofread the manuscript.Competing interests The authors declare no competing interests.Additional information
Supplementary information is available for this paper at https://doi.org/10.1038/s41586-019-
1917-5.
Correspondence and requests for materials should be addressed to Q.W. or X.W.
Peer review information Nature thanks Joseph Loturco, Christopher Walsh and the other,
anonymous, reviewer(s) for their contribution to the peer review of this work.
Reprints and permissions information is available at http://www.nature.com/reprints.