In this study, we sought to measure what changes cells experienced upon microtubule destabilization after inhibiting Rho GTPases. We used human pulmonary microvascular endothelial cells (HPMECs) as a model to measure cell response to microtubule depolymerization in a sensitive cell type. Overall design: RNA-seq profiling of primary human pulmonary microvascular endothelial cells (HPMECs). First, cells were pre-treated with 1 µg/ml cell-permeable C3 transferase (Rho inhibitor, hereafter labeled RHOi) for 4 hours. Following pre-treatment, cells were treated with either 0.1% DMSO or 100 nM vinblastine for 6 hours, while concurrently adding an additional 0.5 µg/ml C3 transferase (RHOi) booster dose. Cells from three donors were purchased from PromoCell. Donor identification between the metadata and data file names corresponds as follows: donor 1 = donor 1, donor 2 = donor 4, donor 3 = donor 6.