Whole cell response to receptor stimulation involves many deep and distributed subcellular biochemical processes

To identify subcellular processes that enable a whole cell response, we stimulated Neuro 2A cells with the cannabinoid receptor agonist HU210 that triggers neurite outgrowth. After 4 different stimulation periods we harvest the cells and subjected them to bulk RNAseq. Differentially expressed genes after each stimulation time were subjected to dynamic and standard enrichment analysis using the Molecular Biology of the Cell Ontology. We predicted the involvement of multiple canonical subcellular processes (SCPs) . Using siRNA ablation experiments of representative SCP genes we documented the involvement of the predicted SCPs in neurite outgrowth. Overall design: Neuro 2A cells were treated with HU210 or drug-free DMSO for 4 different timeperiods, yielding 8 different conditions. 2 to 3 independent experiments of each condition were pooled to generate 8 different samples.