137607.fig.001a
(a)
137607.fig.001b
(b)
137607.fig.001c
(c)
137607.fig.001d
(d)
Figure 1: D3T potentiates AGE-induced cytotoxicity. (a) SH-SY5Y cells were pretreated for 24 hours with 0–50 μM D3T followed by a 24 hr treatment with BSA or AGE-BSA. MTT was performed as indicated in the Materials and Methods section and absorbances were standardized to 0.1% DMSO treatment. Results are from three independent experiments ( ); bar values are expressed as mean ± SD. (b) GSH was measured by HPLC and standardized to total protein. Results are from four independent experiments ( ); bar values are expressed as mean ± SD (* ; ** ; *** ). (c) Cells were pretreated with 0.1% DMSO or 50 μM D3T and then treated with 4.0 mg/mL BSA, AGE-BSA or an equivalent dilution of PBS. Protein oxidation was determined using an OxyBlot kit (Millipore). Band intensity was quanitified by UN-SCAN-IT Gel 6.1 software (Silk Scientific, Orem, UT, USA) and standardized relative to PBS control with DMSO pretreatment. Results are from three independent experiments; bar values are expressed as mean ± SD. (d) Cells were pretreated with 0.1% DMSO- or 50 μM D3T and then treated with 4.0 mg/mL BSA, AGE-BSA ± 2.0 mM NAC, or an equivalent dilution of PBS. Neurite number was quantified using Image J software. Results are from three independent experiments ( random views); values are expressed as mean ± SEM. Data were analyzed by ANOVA. Superscript letters that are not the same are significantly different ( ).