FIGURE 2.

Rin1 depletion from A549 cells resulted in a decrease in proliferative signaling. A, A549 cells were transfected with 10 nmol of either non-targeting control morpholino or Rin1 5′-UTR targeting morpholino and assessed for defects in cell proliferation. B, quantification of the colony formation data is presented. Error bars represent the mean 1 ± S.D., n = 3. C, to assess whether this defect in cell proliferation was because of defects in EGFR signaling, morpholino-treated cells were serum-starved and then stimulated with 100 ng/ml EGF for the indicated times, and cell lysates were analyzed for changes in pERK1/2 levels by Western blot (WB) analysis. D, changes of pERK1/2 levels were measured using densitometry (ImageJ) and normalized to GAPDH levels. Statistical significance of control versus Rin1 depletion was analyzed using a two-way analysis of variance where control versus depletion had a statistical significance at p < 0.001 with F(1,22) = 23.827.