The RT-qPCR and immunoblot analysis of mAb SO57 HC and LC in transgenic tobacco plants.

(A) Relative expression levels (Y axis) of the HC and LC gene as determined by RT-qPCR. Amplicons generated with the RT-qPCR were confirmed by 1% agarose gel electrophoresis (upper panel). The results of RT-qPCR are expressed as the average of three independent experiments after normalization with N. tabacum actin bands (lower panel). Error bars represents mean relative expression values (mean ± SD) of HC and LC to actin from the mAb^P and mAb^PK samples. Transcript levels of HC and LC were not significantly different between the mAb^P and mAb^PK (p>0.05). mAb^P, non-KDEL-tagged mAb^P; mAb^PK, KDEL-tagged mAb^P. (B) Western blot analysis of proteins extracted from leaves of randomly selected 4 transgenic plants without KDEL and 3 transgenic plants with KDEL, respectively. The bands for HC (50 kDa) and LC (25 kDa) were detected with HRP-conjugated goat anti-human Fcγ- or F(ab′)₂-specific antibodies, respectively. *p<0.05 compared to mAb^P samples (Student's t-test analysis). Error bars represent the mean ± SD.