Identification of ALKBH5-interacting RNAs in lung cancer cells by RNA immunoprecipitation sequencing (RIP-Seq)

This experiment was performed to identify the RNAs, particularly lncRNAs, that bind to the m6A demethylase ALKBH5 under nutrient stress. Lysates from 95D cells cultured under glucose- or glutamine-deficient conditions were subjected to immunoprecipitation using an antibody against ALKBH5. Co-precipitated RNAs were isolated, used to construct sequencing libraries, and analyzed to identify enriched transcripts. Overall design: Immunoprecipitation using an anti-ALKBH5 antibody compared to a total RNA input control. 95D cells were cultured under nutrient deprivation for 48 hours to induce the relevant cellular state.