Fig. 1

Simultaneous ribosome profiling/RNA-seq under hypoxic stress a Overview of experiments in ARPE-19 cells subject to hypoxic stress conditions. ARPE-19 cells were prepared in four separate dishes each containing 4 × 10⁸ cells. All time points greater than 0 h were cultured under 1% oxygen for hypoxic stress, and ARPE-19 cells cultured under 21% oxygen served as a control. Cells were harvested and analyzed using western blotting and high though-put sequencing technologies. b HIF-1α and HIF-2α (Epas1) protein levels in ARPE-19 cells were determined by western blotting. c Overall experimental design for simultaneous ribosome profiling and RNA-seq under hypoxic stress. ARPE-19 cells subject to hypoxic stress treatment for 0 h, 1 h, 2 h and 4 h. Polysome RNAs were extracted and digested with nuclease to construct the sequencing library for ribosome profiling, and total RNA was simultaneously extracted for RNA-sequencing (c)