a) Real-time RT-PCR quantification of vspB transcripts in R1 transgenic maize tissue. Two hundred micrograms of total RNA from the indicated tissue was used as the template source for real-time RT-PCR detection of vspB transcripts. Reactions were performed in 15 uL volume using the Qiagen Quantitect SYBR Green kit. Expression values are calculated by normalizing all threshold cycles (Ct) for vspB to the 18S rRNA Ct and converting this value to fold-increase over the value for the lowest expressing tissue, *44-1-1-stem, which was arbitrarily set at 1.0. b,c) Developmental changes in VSPβ level in transgenic maize vegetative tissues. Soybean VSPβ was immunodetected in Western blots of 30 μgs of total protein from crude extracts separated by 12% SDS-PAGE and blotted to Hybond-P membranes. Crude Extracts were prepared from: YL, leaves from preflowering plants; SL, leaves from silage stage plants; and SS, stems from silage stage plants. (b) Coomassie blue stained SDS-PAGE separated proteins from crude extracts; (c) VSP was immunodetected using VSPβ antiserum and the Renascence kit chemiluminescent detection method (NEN Life Sciences Products, Inc). The arrows indicate the VSP-β protein band.