hTCEpi cells were cultured in keratinocyte–serum-free media (K-SFM) with supplements (Invitrogen). Primary HCEKs and HLEKs were isolated from cadaver donor corneas provided by Midwest Eye Bank (Ann Arbor, MI, USA) and cultured in CnT-20 media with supplements (CellnTech, Bern, Switzerland) on collagen IV-coated plates (Corning, Corning, NY, USA). To silence gene expression, small interfering RNAs (siRNAs) (20 nM) targeting ATF3 (Catalog #AM16708; Invitrogen), PBK (Catalog #4390824; siPBK#1: Assay ID s31708, siPBK#2: Assay ID s31707, siPBK#3: Assay ID s31706; Invitrogen) or a scrambled negative control (Catalog #465372; Invitrogen) were transiently transfected into cells using RNAiMAX (Invitrogen). Although we have tested three siRNA sequences for PBK1, only two of them (#1 and #3) reduced the expression efficiently.
Kaplan N., Wang J., Wray B., Patel P., Yang W., Peng H, & Lavker R.M. (2019). Single-Cell RNA Transcriptome Helps Define the Limbal/Corneal Epithelial Stem/Early Transit Amplifying Cells and How Autophagy Affects This Population. Investigative Ophthalmology & Visual Science, 60(10), 3570-3583.
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