Figure 5. Reduction of NAD-IDH enzyme activity by interaction with TBT in an E. coli expression system.

(a) After the expression of pHIDHα, β, γ was induced by culturing E. coli BL21 transformants at 25°C for 20 h, crude extracts were prepared and were subjected to western blot analysis using the anti-NAD-IDHα and anti-GAPDH antibodies. Cropped blots were shown and the full-length blots were indicated in Supplementary Fig. 4. (b) After treatment with 100 nM TBT for 1 h, NAD-IDH activity was determined in the crude extracts. ADP (100 μM) and Zn²⁺ (2 mM) were used as positive and negative controls, respectively. * P < 0.05 compared with the corresponding control group.