Figure 3.

Analysis of the nptII expression in the 271 Lo2 and 271 Lo1 hybrid plants and the segregating progenies. (A) Northern blot analysis of the nptII reporter transgene expression in F1 hybrids. About five micrograms of total RNA were loaded per line and hybridized against the nptII DNA probe. (B) Analysis of the nptII expression by quantitative RT-PCR in the 271 Lo2 and 271 Lo1 hybrids over three generations (F1–F3). The nptII RNA levels were normalized to tobacco actin mRNA. The expression level of the non-silenced HeLo2 parental line has been arbitrary chosen as 100%. Three to five plants of each line were analyzed in three technical replicates. (C) Analysis of the nptII expression by quantitative RT-PCR in the progenies that inherited locus 2 (left) and locus 1 (right) targets without the 271 silencer. S1, S2: first generation and second generation of segregants, respectively.