Quantification of the effects of LRRK2i and AnnexinV on Tat induced neuronal axon elimination, neuronal axon length, and microglia process length. (A) Approximately 80% of axons remained in microfluidic chambers after exposure to saline, DMSO, or LRRK2i alone treated BV-2 cells, where only 20% of axons remained after the addition of Tat treated BV-2 cells. The addition of LRRK2i or AnnexinV to Tat treated BV-2 cells significantly increased survival rate to 63% and 75%, respectively (n = 3 chambers per condition, *P <0.05, **P <0.01, one way ANOVA, Newman-Keuls post-test). (B) Axons exposed to Tat-treated BV-2 cells are significantly shorter post treatment. Axons exposed to either DMSO, LRRK2i alone, Tat and LRRK2i, or Tat and AnnexinV treated BV-2 cells exhibited no significant change. Axons exposed to saline treated BV-2 cells were significantly longer post treatment (n = a minimum of 30 axons per experimental condition, ***P <0.001, two way ANOVA, Bonferroni post test). (C) Tat treated microglia had significantly longer processes than saline treated microglia. LRRK2 kinase inhibition normalized the microglia process length in the presence of Tat. BV-2 cells exposed to Tat and AnnexinV were not significantly different from Tat alone (n = longest process of 100 microglia in co-culture, *P <0.05, **P <0.01, ***P <0.001, one way ANOVA, Newman-Keuls post-test). ANOVA, analysis of variance; DMSO dimethyl sulfoxide; LRRK2i, leucine-rich repeat kinase 2 inhibitor; Tat, trans activator of transcription.