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PC12 clones using a glucose transport and luciferase-based bioluminescent assay as described in MATERIALS AND METHODS. The rate of glucose uptake and amounts of ATP in untreated PC12-VT were set to 100% (mean ± S.E., three independent experiments. *P<0.05, #P<0.01; comparison between the indicated clones. Essentially similar results were obtained using an additional stably transfected UCP4 clone. (D) The indicated View Within ArticlePC12 clones were switched to serum-free medium for 6 h and exposed to vehicle (Veh), NGF (50 ng /mL), EGF (50 ng /mL) either alone or in combination with 3-nitropropionic acid (3-NP) for an additional 24 h. Cell viability was determined using LDH assay as described in MATERIALS AND METHODS. Values represent means±SE of total LDH release from 3experiments each performed in triplicate. *P0.05,#P<0.01; comparison between the indicated clones. Essentially similar results were obtained using an additional stably transfected UCP4 clone.

Supplemental Figure 1. (A) Treatment with 20 μmol/L BAPTA inhibits ERK1/2 activation in primary rat cortical neurons exposed to 20 μmol/L 2,4 dinitrophenol (DNP). Activation of ERK1/2 was determined by Western blotting using phospho-specific and total ERK1/2 antibodies.

Supplemental Figure 2. The indicated PC12 clones were incubated with vehicle (Veh) or 50μmol/L PD98059 (PD) for 1 and 6 h and the ADP/ATP ratio in extracts were determined as described in MATERIALS AND METHODS. The amount of ADP/ATP in untreated PC12-VT was set to 100%. Values represent mean ± S.E., three independent experiments, *P<0.05; #P<0.01. Essentially similar results were obtained using an additional stably transfected UCP4 clone.

Supplemental Figure 3. (A) Level of UCP2 mRNA in the indicated PC12 clones following exposure to 20 μmol/L 3-NP. UCP2 and actin mRNAs were determined by semi-quantitative RT-PCR. (B) ERK1/2 activation in primary rat cortical neurons exposed to either 20 μmol/L DNP or 20 μmol/L 3-nitropropionic acid (3-NP). (C) Pre-treatment with 20 μmol/L DNP maintains ERK1/2 activation in cortical cultures following exposure to 20 μmol/L 3-NP. (D) Pre-treatment with 20 μmol/L DNP maintains ERK1/2 activation in PC12 cells following exposure to 20 μmol/L 3-NP. Activation of ERK1/2 was determined by Western blotting using phospho-specific and total ERK1/2 antibodies.