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. Author manuscript; available in PMC: 2022 Jun 2.
Published in final edited form as: J Neurorestoratology. 2013 Jul 23;1:1–12. doi: 10.2147/jn.s43745

Figure 2.

Figure 2

Lipoic acid promotes hNSC survival in vitro.

Notes: Representative images of TUNEL-positive hNSCs 24 hours after treatment with (A) vehicle (dH2O and ethanol), (B) 400 μM SIN-1 and vehicle (ethanol), and (C) 400 μM SIN-1 following a 24-hour preincubation with 0.25 mM LA. DAPI nuclear counterstain in blue, TUNEL in green, and merged images are shown. Scale bar = 20 μm. (D) Cumulative TUNEL labeling in hNSCs treated with a 24-hour preincubation with LA followed by SIN-1 for 24 hours. Data normalized to the 0 mM LA/0 μM SIN-1 control group (white bar). N = 3/group. **P < 0.05 compared to 0 mM LA/0 μM SIN-1 control. +P < 0.05 +++P < 0.001 compared to 0 mM LA/400 μM SIN-1. Data analyzed by Student’s t-test (E) LDH release from 0.25 mM LA-pretreated hNSCs 24 hours after exposure to SIN-1. Data normalized to 0 μM SIN-1/0 mM LA control. N = 8/group. ***P < 0.001, compared to 0 μM SIN-1/0 mM LA control. +++P < 0.001, compared to associated white bars. Data analyzed by two-way ANOVA with Bonferroni posttests. Serial SIN-1 concentrations in 0 mM LA groups analyzed by one-way ANOVA with Dunnett’s posttest (F) Caspase 3 activity in hNSCs 24 hours after treatment with H2O2, which followed a 24-hour preincubation period with 0.25 mM LA. N = 3/group. ***P < 0.001, compared to 0 μM H2O2/0 mM LA control. +++P < 0.001, compared to associated white bar. Data analyzed by two-way ANOVA with Bonferroni posttests. 0 μM H2O2/0 mM LA and 400 μM H2O2/0 mM LA compared by Student’s t-test (G) WST-1 cell viability assay of hNSCs exposed to hypoxia (3% oxygen) for 24 hours followed by a 4-hour reperfusion at normoxia. Cells were treated with vehicle (ethanol) or 0.25 mM LA for 24 hours prior to hypoxia. Data normalized to the vehicle control group. N = 12/group. ***P < 0.001. Data analyzed by Student’s t-test.

Abbreviations: ANOVA, analysis of variance; DAPI, 4′,6-diamidino-2-phenylindole; hNSCs, human neural stem cells; LA, lipoic acid; min, minute; TUNEL, terminal-deoxynucleotidyl-transferase-mediated dUTP nick end labeling; WST, water-soluble tetrazolium.