Time courses analysis of the effect of latrunculin B-treatment on gene expression in in vitro differentiated pancreatic progenitor cells.

The organization of the actin cytoskeleton is known to influence the endocrine versus ductal fate choice during pancreas development via YAP. However, the intermediate mechanism between changes to actin polymerization and changes in YAP activity during pancreas development has not been identified. Pharmacological inhibition of actin polymerization has also been shown to be a stronger inducer of endocrine fate choice than pharmacological inhibition of YAP activity, indicating that actin depolymerization may induce endocrine fate choice via additional pathways. To identify these mechanisms, we performed bulk RNA-sequencing of in vitro differentiated ESC-derived pancreatic progenitor cells treated with latrunculin B for 3, 6, 12, or 24 hours. This sequencing dataset revealed multiple promising candidates for actin depolymerization-induced YAP inactivators and YAP-independent endocrine induction, which have been validated by additional methods. To investigate the intermediate effects between actin depolymerization and endocrine specification, ESC-derived pancreatic progenitor cell were treated with the actin depolymerization agent latrunculin B for 3, 6, 12 or 24 hours. These samples were compared to samples harvested before treatment and samples harvested after 24 hours of DMSO treatment. Duplicates of 24hr untreated and 12hr LatB treated and triplicates of other samples were sequenced.