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. Author manuscript; available in PMC: 2016 Nov 1.
Published in final edited form as: J Neuropathol Exp Neurol. 2015 Nov;74(11):1093–1118. doi: 10.1097/NEN.0000000000000255

Figure 2.

Figure 2

Antiretrovirals do not alter oligodendrocyte precursor cell (OPC) number or induce apoptotic cell death. (A, B) Primary mouse OPCs plated on coverslips were put into differentiation medium and treated with vehicle (DMSO), doses of Zidovudine (AZT) (1 μM, 10 μM, or 25 μM), Ritonavir (100 nM, 1 μM, or 3 μM), or Lopinavir (150 nM, 1.5 μM, or 15 μM). After 72 hours, cells were fixed and stained with antibody to A2B5 with DAPI (blue) for total cell nuclei. Results from AZT are in the top row, Ritonavir in the center, and Lopinavir in the bottom row respectively. Epifluorescent images were captured, with 10 fields on 3 coverslips per condition. (A) Total cell number (DAPI-positive nuclei) were counted using ImageJ software from the biological replicates utilized for the dose curves in Figures 1 and 2 (n = 8). Results are represented as percentage normalized to untreated. Significance compared with control (#p < 0.05). Data are presented as mean ± SE. One-way ANOVA followed by post-hoc Newman-Keuls determined statistical significance. (B) Oligodendrocyte precursor cells (A2B5-positive cells) were counted using ImageJ software and are represented as percentage normalized to untreated. Data are presented as mean ± SE. Results from n = 3 independently prepared cultures. One-way ANOVA followed by post-hoc Newman-Keuls determined statistical significance. (C) Primary mouse OPCs plated on coverslips were put into differentiation medium and treated with vehicle (DMSO) or doses of Lopinavir (1.5 μM, or 15 μM). After 72 hours, cells were fixed and stained using a TUNEL assay to label cells with double-stranded DNA breaks indicative of activation of the apoptotic cascade. DNase was used as a positive control. Epifluorescent images were captured at 40x magnification, with 10 fields on 3 coverslips per condition. Results from n= 2 independently prepared cultures. Data are presented as mean ± SE. One-way ANOVA followed by post-hoc Dunnett’s comparison determined statistical significance. Significance compared with control (###p < 0.001).