Reagent or resource	Source	Identifier
Antibodies
Sheep anti‐ChAT	ABCAM	ab18736; RRID:AB_2244867
Goat anti‐ChAT	Millipore	AB‐144P; RRID:AB_2079751
Rabbit anti‐EN1 86/8	Home made	Sonnier et al (2007)
Rabbit anti‐EN1 LSBio	CliniSciences	LS‐B9070
Mouse anti‐neurofilament 165kDa	DSHB	2H3; RRID:AB_531793
Mouse anti‐SV2A	DSHB	SV2; RRID:AB_2315387
Rabbit anti‐p62/SQSTM1	ABCAM	ab109012; RRID:AB_2810880
Alpha bungarotoxin, Alexa fluor 488 conjugate	Thermo Fisher	B13422
Chemicals
PBS	Thermo Fisher	14200075
Triton X‐100	Sigma‐ Aldrich	9036‐19‐5
Normal Goat Serum	Thermo Fisher	31873
Fetal Bovine Serum	Gibco	26140087
DAPI Fluoromount‐G	Southern Biotech	0100‐20
O‐Xylene	CARLO‐HERBA	403926
Cresyl Violet acetate	Sigma Aldrich	105235
Eukitt Quick‐hardening mounting medium	Sigma Aldrich	03989
Sucrose	Sigma Aldrich	84097
SYBR‐Green	Roche	04887352001
EDTA‐free Complete Proteases inhibitor cocktail	Roche	11873580001
250U/ml of benzonase endonuclease	Sigma Aldrich	E1014‐25kU
Nu‐PAGE 4‐12% acrylamide 1 mm SDS‐PAGE	Invitrogen	NP0321BOX
LDS‐DTT Sample Buffer	Invitrogen	NP0007
Tween‐20	Sigma Aldrich	9005‐64‐5
Commercial assays
RNAscope Multiplex Fluorescent V2 Assay	ACD	323100
RNA‐Protein Co‐detection Ancillary Kit	ACD	323180
RNA Extraction RNeasy Kit	QIAGEN	74104
Chemiluminescence substrate CLARITY ECL	BIORAD	170‐5061
Pierce BCA assay		23225
Oligonucleotides
qPCR Engrailed‐1 sense	This study	CCTGGGTCTACTGCACACG
qPCR Engrailed‐1 antisens	This study	CGCTTGTTTTGGAACCAGAT
qPCR Gapdh sense	This study	TGACGTGCCGCCTGGAGAAAC
qPCR Gapdh antisense	This study	CCGGCATCGAAGGTGGAAGAG
RNAscope Mm‐En1	ACD	442651
RNAscope Mm‐ChAT	ACD	408731‐C2
RNAscope Mm‐Calb1	ACD	428431‐C3
Viruses
scFvMUT	Vector Biolabs	AAV8‐IgK‐scFvMUT‐6xMyc‐2A‐eGFP
scFvEN1	Vector Biolabs	AAV8‐IgK‐scFvEN1‐6xMyc‐2A‐eGFP
Experimental models
En1 lacZ (En1‐Heterozygous; M. musculus)	Jackson Lab	00712
Software
Image Studio Lite	Li‐Cor	https://www.licor.com/bio/image-studio-lite/
GraphPad Prism	GraphPad	https://www.graphpad.com
Fiji	ImageJ	https://imagej.net/Fiji
Other
Grip Strength Meter	AHLBORN	ALMEMO 2450