Figure 5

miR-3127-5p regulates c-Abl/Ras/ERK pathway activation by targeting oncogene c-Abl.

(A): A549 or H292 cells transduced with Lv-3127, Lv-3127-off, or Lv-GFP were lysed. Lysates (1.0 mg) were incubated with 50 μL of GST-Raf1-RBD beads for 30 min to pull down active Ras. Samples were immunoblotted using an anti-K-Ras antibody, thereby allowing levels of GTP-bound Ras within the cells to be assessed. Whole cell lysates (50 µg) were also subjected to western blot for detection of total K-Ras. The numbers under bands indicate the ratio of integrated optical density to GAPDH normalized by GFP control (The full-length blots were presented in the supplementary Figure S5). (B): A549 or H292 cells transduced with Lv-3127, Lv-3127-off, or Lv-GFP were subjected to immunoblotting with antibodies to ERK, p-ERK and downstream factors MMP-9, VEGF and cyclin D1 (The full-length blots were presented in the supplementary Figure S4, 5). (C): Schematic diagram illustrating the effect of reduced miR-3127-5p levels on activation of c-Abl/Ras/ERK pathway and promotion of tumor proliferation and metastasis.