Figure 5.

Functional effects of CDH1 and CADM1 knockdown on MP38 cells: Migration of MP38 cells after treatment with CDH1 and CADM1 siRNAs for 72 hours (A). Cells were trypsinized and subjected to Boyden chamber-based, serum-directed migration assays. Quantification of migrated cells and representative images are shown. Scale bars, 250 mmol/L. Data are graphed as fold change in cell migration compared with untreated cells from at least five independent experiments, P < 0.05, and P < 0.01 as determined by t test, and error bars are SEM. B, Effect of siCDH1 and siCADM1 knockdown on cell adhesion. Percent cell adhesion is calculated as (adherent cell fluorescence)/(total cell fluorescence) from four independent experiments, P < 0.05, and P < 0.01 as determined by t test, and error bars are SEM. C, Cell viability as measured by ATP luminescence (Cell Titer Glo) was studied after siCDH1 and siCADM1. Cells were treated with CDH1 and CADM1 siRNAs for 72 hours, trypsinized, and cultured for 72 hours in low attachment conditions. Fold change compared with control was calculated for each replicate, P < 0.05, and P < 0.01 as determined by t test, and error bars are SEM. Data were collected from three independent experiments. D, Effect of siCDH1 and siCADM1 on spheroid size and cluster formation after being cultured on low attachment conditions for 72 hours. Representative microscopy images of MP38 spheroids from six independent biological replicates were taken after 72 hours. Quantitation of spheroid size was determined by Image J, P < 0.05, and P < 0.01 as determined by t test, and error bars are SEM. Scale bars, 500mmol/L.