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Decreased Tumor Progression and Invasion by a Novel Anti-Cell Motility Target for Human Colorectal Carcinoma Cells

Figure 3

km23-1 regulates the paracrine effects of active TGFβ1 on NIH3T3 cell migration and mitogenesis.

A: NIH3T3 fibroblasts were plated onto polycarbonate membrane filter inserts (8.0 µm pore size) in 6-well Transwells and CM from RKO cells and their siRNA stable transfectants were added to the NIH3T3 cells as described in “Materials and methods.” Cells migrating into the lower chambers were counted. SM: supplemented McCoy's (SM) medium; EV: CM from RKO-EV cells; NC siRNA: CM from NC siRNA-RKO cells; km23-1-siRNA clone #1: CM from km23-1-siRNA-RKO clone #1; km23-1-siRNA clone #5: CM from km23-1-siRNA-RKO clone #5; NC siRNA/Tβ1Ab: CM from NC-siRNA cells that had been incubated with a neutralizing anti-TGFβ1 antibody (30 µg/ml); Ctrl/IgG: CM from Ctrl with IgG. Data plotted are the mean ±SE of triplicate wells from a representative experiment (n = 3). *p<0.01 compared to either NC-siRNA or NC-siRNA/IgG. B: NIH3T3 cells were plated in the lower chamber and made quiescent as described in the “Materials and methods.” RKO cells and their siRNA stable transfectants were then plated onto polyester membrane filter inserts (0.4 µm pore size) in 12-well Transwells. Mitogenesis assays were performed as described in “Materials and methods” to assess the effect on NIH3T3 cell mitogenesis after co-culture with RKO stable transfectants. Data plotted are the mean ±SE of triplicate wells from a representative experiment (n = 3). *p<0.01 compared to the NC-siRNA or NC-siRNA/IgG.

Figure 3

doi: https://doi.org/10.1371/journal.pone.0066439.g003