Expression and effects of MIA in cultured pancreatic cancer cells. A: MIA mRNA levels in indicated pancreatic cancer cell lines were determined by real-time quantitative polymerase chain reaction, as described in the Methods section. Data are presented as median + SD of MIA mRNA copies per μl of input cDNA normalized to housekeeping genes CPB and HPRT. B: 30 μg protein lysates of the indicated cell lines were subjected to immunoblotting analysis using a specific MIA antibody, as described in the Methods section. C: Immunoprecipitation analysis was carried out as described in the Methods section. D: An in vitro cell invasion assay was performed using 8 μM filters coated with Matrigel, as described in the Methods section. 4 × 105of the indicated pancreatic cancer cells were seeded onto the filters in 10% serum overnight, and then treated as indicated for 24 h. Invaded cells were stained and counted. The values shown are the mean ± SEM obtained from three independent experiments.