Figure 2

Binding Affinity and biological effects of PLZ4 against canine TCC cell lines. A. Binding affinity of PLZ4 against K9TCC-PU and K9TCC-PU-In. Twenty thousands cells of K9TCC-PU and K9TCC-PU-In were seeded in 96 well plates. After culture for 24 hours, cells were fixed and incubated with different concentrations of PLZ4-biotin for 1.5 hours followed by SA-HRP for another 1 hour. Cells treated with SA-HRP alone served as background control. The color was developed using TMB substrate and read by ELISA readers. Three independent experiments conducted in triplicate were performed. The mean values of the 3 experiments are shown. B. Biological effects of PLZ4 on canine TCC cell lines. Ten thousand cells of K9TCC-PU-In or K9TCC-PU were seeded in the 96-well plates and treated with increasing concentrations of PLZ4 or PBS for 2 days. The cell proliferation assay was assessed by the WST-8 assay per manufacturer's protocol. Cells treated with PBS were used as 100% control. Each experiment was performed three times in triplicate. Mean values at each concentration are presented.