RNA Sequencing Analysis of H1299 cells with Hyperoxia Treatment and Control Cells

Purpose: To study the effect of hyperoxia on lung cancer cells, H1299 cells were cultured in 60% O2 for 12 h and 24 h, and RNA-seq was performed. Overall design: Total RNAs from the H1299 cells treated with hyperoxia treatment (24 h) and control cells were isolated and quantified. The high-throughput RNA sequencing analysis was performed by Shanghai Aksomics Biotech Company Ltd. (Shanghai, China). Briefly, The ribosomal RNA (rRNA) was removed from the total RNA using the NEBNext® rRNA Depletion Kit (New England Biolabs, Inc., Ipswich, MA, USA) following the manufacturer's instructions. RNA libraries were constructed by using NEBNext® Ultra™ II Directional RNA Library Prep Kit (New England Biolabs, Inc., Massachusetts, USA) according to the manufacturer's instructions. Then, HTSeq software (v0.9.1) was used to obtain the transcript level (lncRNA) raw count as the expression profiling, and edgeR (v3.16.5) was used to perform normalization, and differentially expressed mRNAs were identified by p-value and fold change.