Illumina NovaSeq 6000 sequencing; Transcriptome and secretome analysis of mechanically stretched osteocytic cells reveals activation of signalling pathways implicated in bone regeneration and DNA-repair systems
Transcriptome and secretome analysis of mechanically stretched osteocytic cells reveals activation of signalling pathways implicated in bone regeneration and DNA-repair systems
Immediately after the mechanical loading, cells were washed twice with ice-cold PBS and lysed with RLT buffer (Qiagen) for 1 minute.  RNA was extracted using the RNeasy kit (Qiagen) according to the manufacturer instructions. RNA quantity and quality were estimated using the NanoDrop™ 2000 spectrophotometer (Thermo Scientific™)  Sequencing libraries were generated using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA) following the manufacturer's recommendations and index codes were added to attribute sequences to each sample.