Reagent	Concentration	Source	Identifier
Animal studies
C57BL/6 mice	N/A	The Jackson Laboratory	Stock #000664
Math1-Cre mice	N/A	The Jackson Laboratory	Stock #011104
SmoM2-eYFPloxP/loxP mice	N/A	The Jackson Laboratory	Stock #005130
PhamloxP/loxP mice	N/A	The Jackson Laboratory	Stock #018385
Gli-luc mice	N/A	Generously shared by Dr. Oren Becher, Northwestern University and Dr. Eric Holland, Fred Hutchinson Cancer Research Center	MGI #4820828
N-Methyl-2-pyrrolidone (NMP)	1:10	Millipore Sigma	Catalog #328634
Polyethylene glycol (PEG)	9:10	Millipore Sigma	Catalog #P3015-250G
Vismodegib	75 mg/kg or 100 mg/kg in NMP:PEG	LC Laboratories	Catalog #V-4050
Isoflurane	Vapor	Piramal Critical Care, Inc.	NCD Code #66794-017-25
PCR
Tail lysis buffer	1×	Allele Biotechnology	Catalog #ABP-PP-MT01
Cre forward primer: GCG GTC TGG CAG TAA AAA CTA TC	200 µM	Invitrogen	JAX #oIMR1084
Cre reverse primer: GTG AAA CAG CAT TGC TGT CAC TT	200 µM	Invitrogen	JAX #oIMR1085
SmoM2 (YFP) forward primer: AAG TTC ATC TGC ACC ACC G	400 µM	Invitrogen	JAX #oIMR0872
SmoM2 (YFP) reverse primer: TCC TTG AAG AAG ATG GTG CG	400 µM	Invitrogen	JAX #oIMR1416
Pham Common primer: CCA AAG TCG CTC TGA GTT GTT ATC	200 µM	Invitrogen	JAX #13840
Pham WT reverse primer: GAG CGG GAG AAA TGG ATA TG	200 µM	Invitrogen	JAX #13841
Pham Mutant reverse primer: TCA ATG GGC GGG GGT CGT T	200 µM	Invitrogen	JAX #oIMR7320
Gli-luc forward primer: TATCATGGATTCTAAAACGG	200 µM	Invitrogen	N/A
Gli-luc reverse primer: CAGCTCTTCTTCAAATCTATAC	200 µM	Invitrogen	N/A
Apex Taq DNA Polymerase Master Mix	1×	Genessee Scientific	Catalog #42-138
Platinum Blue PCR SuperMix	1×	Invitrogen	Catalog # 12580015
Molecular biology grade water	–	Corning Inc.	46-000-CM
Immunofluorescence and immunohistochemistry
Paraformaldehyde (PFA)	4% in PBS
Phosphate-buffered saline (PBS)	1×
Antigen retrieval	1:100	Vector Laboratories	Catalog #H-3300
Donkey serum	2% in 0.3% PBST	Millipore Sigma	Catalog #D9663
DAPI	1:2500	Invitrogen	Catalog #D1306
SOX2	1:200	Cell Signaling Technology	Catalog #4900
HES1	1:200	Cell Signaling Technology	Catalog #11988
MyoD1	1:100 (Mouse)	Novus	Catalog #NBP2-32882-0.1 mg
phospho-RB (Ser807/811)	1:3000	Cell Signaling Technology	Catalog #8516
GFAP	1:2000	Dako	Catalog #Z0334
NeuN	1:10,000	Millipore	Catalog #MAB377
Calbindin	1:400	Cell Signaling Technology	Catalog #2173
Goat anti-rabbit Alexa Fluor 488	1:400	Thermo Fisher Scientific	Catalog #A-11034
Goat anti-mouse Alexa Fluor 555	1:400	Thermo Fisher Scientific	Catalog #A-21424
Novolink Polymer	Per manufacturer’s instructions	Leica Biosystems	Catalog #RE7200-CE
ImmPRESS™ HRP Anti-Rabbit IgG	Per manufacturer’s instructions	Vector Laboratories	Catalog #MP-7401
ImmPRESS™ HRP Anti-Mouse IgG	Per manufacturer’s instructions	Vector Laboratories	Catalog #MP-7402
Dissociation
Papain Dissociation System	Per manufacturer’s instructions	Worthington Biochemical Corporation	Catalog #LK003150
Hank’s Balanced Salt Solution (HBSS)	1×	Gibco	Catalog #14175-095
d-(+)-Glucose	6 g/L	Millipore Sigma	Catalog #G7021
Flow cytometry
FIX & PERM Cell Fixation & Cell Permeabilization Kit	Per manufacturer’s instructions	Thermo Fisher Scientific	Catalog #GAS003
Heat-inactivated Fetal Bovine Serum (HI-FBS)	5% in FACS wash buffer	Gibco	Catalog #10437028
Sodium azide (NaN3)	0.1% in FACS wash buffer	Fisher Scientific	Catalog #S2271-25
PFA	0.1% in sheath fluid
FxCycle Violet	1:100	Thermo Fisher Scientific	Catalog #F10347
phospho-RB Alexa Fluor 488	1:50	Cell Signaling Technology	Catalog #4277
MyoD Allophycocyanin (APC)	1:50	Novus	Catalog #NBP2-34772APC
GFAP Alexa Fluor 647	1:50	Cell Signaling Technology	Catalog #3657
Commercial assays
Drop-seq beads32	90–110/μL	ChemGenes	Lot # 090316
Nextera XT	NA	Illumina	FC-131-1024
Oligonucleotides
Template_Switch_Oligo: AAGCAGTGGTATCAACGCAGAGTGAATrGrGrG		32	N/A
TSO_PCR: AAGCAGTGGTATCAACGCAGAGT		32	N/A
P5-TSO_Hybrid:  AATGATACGGCGACCACCGAGATCTACACGCCTGTCCGCGGAAGCAGTGGTATCAACGCAGAGT*A*C		32	N/A
Read1CustomSeqB: GCCTGTCCGCGGAAGCAGTGGTATCAACGCAGAGTAC		32	N/A