Ribo-seq analysis of SHSY-5Y neuronal differentiation model

To understand the translational changes occuring during neuronal differentiation and maturation of SHSY-5Y cells, Ribosome profiling (Ribo-seq) was conducted. First, Differentiated and undifferentiated cells were collected in polysome buffer containing cycloheximide. Next, ribosome foot printing was done using RNAse If. Ribosome protected fragments were then extracted using TBE-UREA gel, end-repaired using T4-PNK, and DNA library synthesis performed using NEBNEXT small RNA Library Preparation Kit.