GSM9229810: shEno_2; Homo sapiens; RNA-Seq
To silence ENO1, cell lines were infected by lentivirus containing short hairpin RNA (shRNA) targeting ENO1 (Obio Technology, China;). After two days, cell lines stably expressing shENO1 (ENO1-knockdown) were enriched by a 2-day puromycin (Gibco, USA) treatment for selecting positive clones. To achieve transient overexpression of ENO1, cells were transfected with the pRLenti-CMV-ENO1-3FLAG-PGK-Puro plasmid (Obio Technology, China) using the Nucleofector Electroporation System (Lonza, Switzerland). Additionally, a blank pRLenti-CMV-MCS-3FLAG-PGK-Puro plasmid was transfected as a negative control. TRIzol reagent was used to extract total RNA and purify it to ensure that the quality and concentration of RNA samples met the requirements of subsequent sequencing. Low-quality sequences, such as head sequence, sequence with mass score <20, original reads with N-base ratio >10% and sequence less than 25bp, were removed from the RNA sequencing results.