Figure 5.

Ago2 reduces cell adhesion molecule expression and homotypic aggregation. A, Bright-field (top panel) and fluorescent Calcein-AM (bottom panel) images of fixed cell aggregates from a representative experiment are shown. Quantification of the homotypic aggregation assay was obtained from three independent experiments performed in duplicate, and values under each picture represent the average number of aggregates per ml of sample + sem. Scale bars represent a length of approximately 350 μm. B, Total RNA from the cell types mentioned previously was analyzed via real-time PCR for β-actin, E-cadherin, vinculin, β-catenin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA levels. Values were normalized to RPL-19 and reported as the mean + sem, relative to the MCF-7-Par samples. C, Western blot analyses on the cell types noted previously. Each blot indicates the representative expression of β-actin, E-cadherin, β-catenin, and tubulin in each of the respective cell types. Values under each band represent the average densitometric intensity of protein expression relative to the MCF-7-Par cell line. Data collected from the Ago2 transfectants are representative of three stably selected pools of cells with equal WT or mutant Ago2 expression (*, P < 0.05, compared with MCF-7-Par control).