To investigate which cytokines may induce Rab7a activation in ischemic stroke, we cultured primary mouse BECs in vitro under conditions of either oxygen and glucose deprivation (OGD), or inflammation (TNFα and IL-1β; 10 ng/mL) for 48 h and quantified the amount of GTP-bound Rab7a by using a GST-RILP fusion protein to pull down selectively the active protein from cell lysates as described [31, 33] (Fig. 7A; see Methods for more details). The gene discussed is TNF; the disease is ischemic stroke.