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Determination of Free and Bound SA The third true leaves (counted from below) of 6070 days-old tobacco plants were analyzed to determine free and bound forms of SA essentially as described by Analyses of TMV Accumulation and Tobacco Defense Gene Expression To analyze TMV accumulation and tobacco defense gene expression 200 mg fresh plant leaf tissue per sample was collected and grounded in liquid nitrogen

The cells were then collected by centrifugation, washed with cold PBS and incubated with annexinV-Pacific Blue in annexin-binding buffer at 20 C for 15 min

The hemolytic process may be fully compensated, although mild to moderate anemia is the rule (hemoglobin 8 to 10 g/dl)

Glutathione is an important antioxidant that helps protect against oxidative stress