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Fig. 3 | BMC Research Notes

Fig. 3

From: Production of a mono-biotinylated EGFR nanobody in the E. coli periplasm using the pET22b vector

Fig. 3

Effect of monovalent or tetravalent EL2BH on PC9 cells. a After inoculation of PC9 cells with monovalent EL2BH nanobody, streptavidin (SA), tetravalent EL2BH (EL2BH + SA) or PBS as vehicle control, cells were incubated for 24 h in low serum condition (Opti-MEM), and cell lysates were prepared. Proteins were size-separated by SDS-PAGE and blotted onto PVDF membranes. Membranes were then stained with antibodies against phosphorylated EGFR (Tyr1068), pAKT (Ser473), pERK (Tyr185/187), and for actin as loading control. b PC9 cells were seeded in a 96 well plate 24 h prior to inoculation as in a and cell confluency was further observed in an IncuCyte® live cell analyzer for 8 days. The differences between groups were calculated by comparing the area under the curve (AUC) of each phase confluency graph from 4 different experiments, using ANOVA followed by the Tukey test. The difference between groups was considered statistically significant when P < 0.05*, P < 0.01** and P < 0.001***. c Representative images from b on day 8

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