HiSeq X Ten paired end sequencing; RNA-seq to investigate the effect of curcumin on hypoxia-induced proliferation of human retinal pigment epithelium ARPE-19 cells
RNA-seq to investigate the effect of curcumin on hypoxia-induced proliferation of human retinal pigment epithelium ARPE-19 cells
10^7 ARPE-19 RPE cells treated with 100 μM CoCl4 and 15 μM curcumin were harvested for RNA-seq Total RNA was isolated from the samples with RNeasy Mini kit (QIAGEN, Gemany) in accordance to the manufacturers protocol. RNA was quantified using the NanoPhotometer spectrophotometer (Implen Inc., CA, USA). We then measured RNA concentration using Qubit RNA Assay Kit in Qubit 2.0 Flurometer (Life Technologies, CA, USA), and assessed RNA integrity using the RNA Nano 6000 Assay Kit in Bioanalyzer 2100 (Agilent Technologies, CA, USA). All the samples had a high RNA Integrity Number (RIN) score (>9.4). A total of 1ug of RNA per sample was submitted to the Genefund Co. Ltd (Shanghai, China) for library generation and sequencing.