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. 2016 Dec 15;14(12):e1002583. doi: 10.1371/journal.pbio.1002583

Fig 2. Unbiased screening identifies chemically diverse candidate protective agents that reduce Psy toxicities.

Fig 2

(A) Schematic depicting the work flow for unbiased screening with Celigo adherent cytometer (Nexcelom). (B) Representative plot of relative cell number over 5 d for rat O-2A/OPCs exposed to 1 μM Psy or vehicle (DMSO). Quantification of the relative proliferation rate for all 1,040 drugs at three concentrations over 5 d. Blue: “hit” drugs selected for further validation. A bar graph quantifying the relative proliferation rate of all vehicle and Psy controls is included. (C) Quantification of the relative proliferation rate of rat O-2A/OPCs exposed to 1.5 μM Psy, with and without verified pro-division “hits,” over 5 d. (D) Quantification as in (C) for cells exposed to Psy 48 h before exposure to the indicated drugs. (E) Summary of clinical metadata for lead “hits” as % of total. (* worldwide approval, including FDA. ** reported blood–brain barrier permeability; not all drugs have been examined/reported.) (F) Quantification of cell survival in cells exposed to 3.3 μM Psy for 5 d, with and without the indicated drugs, for 5 d. (G, H) Quantification of the (G) relative survival and (H) number of rat O-2A/OPCs per clone exposed to of Psy (1 μM in H), with and without 100 ng/mL insulin-like growth factor (IGF-1), for 5 d. (I–K) Quantification of (I) neutral lipid and phospholipid accumulation, (J) cathepsin B and D activities in rat O-2A/OPCs exposed to the indicated drugs, with and without 1 μM Psy, and (K) time to half-maximal intensity for endocytosis of fluorescent nanobeads for (I) 2 d or (J, K) 24 h. (L) Quantification of lysosomal pH for rat O-2A/OPCs exposed to 1 μM Psy, with and without the indicated “hits” (blue) or “non-hits” (gray), for 24 h. NT-3: 10 ng/mL; 1E04: 5 μM; 5F05, 9A06, 9H10: 1 μM. ap < 0.05, bp < 0.01 versus Psy-only. Data for all graphs displayed as mean ± SEM; ns = not significant; *p < 0.05, **p < 0.01, †p < 0.001 versus untreated; ap < 0.05, bp < 0.01, cp < 0.001 versus Psy-only treatment. See also S2 Fig, S1 and S2 Tables for drug names and concentrations. Data presented in this figure can be found in S1 Data.