Growth Protocol:	Human Skeletal Muscle Myoblasts (HSMM) derived from quadricep biopsy (Lonza; catalog CC-2580; lot 257130) were expanded in 10% FBS SkGM-2 (Lonza). All procedures have been performed using HSMM within P10 from explant.
Treatment Protocol:	HSMM cell were differentiated for the via switch to 2% HS aMEM (Lifetech).; HSMM cells were differentiated for the via switch to 2% HS aMEM (Lifetech).
Extract Protocol:	[TruSeq] - HSMM in either growth or differentiation medium were dissociated, washed and resuspended in complete media containing 0.1U/ul of RNaseOUT (Lifetech). Each time point was collected in three independent biological replicates for regular mRNA sequencing while for single cell mRNA sequencing the independent replicates were pooled in equal amounts. For bulk RNA-Seq libraries, total RNA was extracted with Trizol and mRNA libraries were produced starting from 100ng of total RNA using the TruSeq mRNA-Seq library kit (Illumina) according to manufacturer’s instructions.; For single-cell mRNA sequencing, dissociated cells were captured and processed with the C1 Single-Cell Auto Prep System (Fluidigm) following manufacturer’s protocol 100-5950. Starting with a suspension of cells at a concentration of approximately 250 cells/ L, up to 96 single cells are captured in each C1 microfluidic device.
Library Construction Protocol:	For bulk RNA-Seq libraries, total RNA was extracted with Trizol and mRNA libraries were produced starting from 100ng of total RNA using the TruSeq mRNA-Seq library kit (Illumina) according to manufacturer’s instructions.; [TruSeq] - HSMM in either growth or differentiation medium were dissociated, washed and resuspended in complete media containing 0.1U/ul of RNaseOUT (Lifetech). Each time point was collected in three independent biological replicates for regular mRNA sequencing while for single cell mRNA sequencing the independent replicates were pooled in equal amounts. For bulk RNA-Seq libraries, total RNA was extracted with Trizol and mRNA libraries were produced starting from 100ng of total RNA using the TruSeq mRNA-Seq library kit (Illumina) according to manufacturer’s instructions. [Single Cell] - After imaging with a microscope to identify which sites have captured a single cell, processing of the cells occurs within the C1 instrument to perform the steps of cell lysis, cDNA synthesis with reverse transcriptase, and PCR amplification of each cDNA library. The cDNA synthesis and PCR use reagents from the SMARTer Ultra Low RNA Kit for Illumina Sequencing (Clontech 634936). The SMARTer chemistry utilizes a strand-switching mechanism so that both the 1st and 2nd strands of cDNA are synthesized in a single reaction. Following harvest from the C1 microfluidic device, each cDNA library is subjected to tagmentation (simultaneous fragmentation and tagging with sequencing adapters) using the Nextera XT DNA Sample Preparation Kit (Illumina FC-131-1096) as described in Fluidigm protocol 100-5950. PCR amplification of the tagmented cDNA uses Index Primers from the Nextera XT DNA Sample Preparation Index Kit (Illumina FC-131-1002). Following PCR, the cDNA libraries from individual cells are pooled and purified using AMPure XP beads (Agencourt Bioscience Corp A63880) as described in Fluidigm protocol 100-5950.; [Single Cell] - After imaging with a microscope to identify which sites have captured a single cell, processing of the cells occurs within the C1 instrument to perform the steps of cell lysis, cDNA synthesis with reverse transcriptase, and PCR amplification of each cDNA library. The cDNA synthesis and PCR use reagents from the SMARTer Ultra Low RNA Kit for Illumina Sequencing (Clontech 634936). The SMARTer chemistry utilizes a strand-switching mechanism so that both the 1st and 2nd strands of cDNA are synthesized in a single reaction. Following harvest from the C1 microfluidic device, each cDNA library is subjected to tagmentation (simultaneous fragmentation and tagging with sequencing adapters) using the Nextera XT DNA Sample Preparation Kit (Illumina FC-131-1096) as described in Fluidigm protocol 100-5950. PCR amplification of the tagmented cDNA uses Index Primers from the Nextera XT DNA Sample Preparation Index Kit (Illumina FC-131-1002). Following PCR, the cDNA libraries from individual cells are pooled and purified using AMPure XP beads (Agencourt Bioscience Corp A63880) as described in Fluidigm protocol 100-5950.

Molecule Type:	poly(A)+ RNA
Library Source:
Library Layout:	PAIRED
Library Strand:	-; Forward
Platform:	ILLUMINA
Instrument Model:	Illumina HiSeq 2500; Illumina HiSeq 2000
Strand-Specific:	Unspecific; Specific

Data Resource	GEN Sample ID	GEN Dataset ID	Project ID	BioProject ID	Sample ID	Sample Name	BioSample ID	Sample Accession	Experiment Accession	Release Date	Submission Date	Update Date	Species	Race	Ethnicity	Age	Age Unit	Gender	Source Name	Tissue	Cell Type	Cell Subtype	Cell Line	Disease	Disease State	Development Stage	Mutation	Phenotype	Case Detail	Control Detail	Growth Protocol	Treatment Protocol	Extract Protocol	Library Construction Protocol	Molecule Type	Library Layout	Strand-Specific	Library Strand	Spike-In	Strategy	Platform	Instrument Model	Cell Number	Reads Number	Gbases	AvgSpotLen1	AvgSpotLen2	Uniq Mapping Rate	Multiple Mapping Rate	Coverage Rate