RNA seq for HLECs transfected with siID3
To understand the role of ID3 in human limbal epithelial cells (HLECs), HLECs were transfected with siID3 or sicontrol . RNAs isolated from these cells were subjected to RNA seq analysis Overall design: HLECs from 3 donors were transfected with a 20 nM siRNA pool against ID3 (Invitrogen, Carlsbad, CA, USA) or nontarget control (Invitrogen, Carlsbad, CA, USA) as previously described for 3 days. Then, cells were processed for total RNA isolation using miRNeasy kit (Qiagen, Hilden, Germany). Expression profiling was conducted using a Next-Generation Sequencing platform at Northwestern University Center for Genetic Medicine core. Gene differential expression profiles obtained from RNA-seq analysis were subjected to gene ontology analysis using DAVID.