This RNA-seq dataset was generated from day 11 differentiated myocytes derived from an immortalized FSHD1_76 cell line without electrical pulse stimulation and represents replicate 3 of the experiment. Total RNA was extracted by using the RNeasy kit (74106, QIAGEN). RNA was converted to cDNA using the Smart-Seq 2 protocol. DNA libraries were constructed using Nextera DNA Flex Library Prep Kit (Illumina). DNA samples were sequenced on the Illumina NovaSeq6000 platform with PE100 to a depth of around 20 million reads per sample.