Journal: The Journal of Biological Chemistry

Article Title: Dicer Knockdown Inhibits Endothelial Cell Tumor Growth via MicroRNA 21a-3p Targeting of Nox-4 *

doi: 10.1074/jbc.M113.519264

Figure Lengend Snippet: MicroRNA biogenesis is essential for endothelial cell tumor formation. EOMA cell stable transfectants were generated using lentiviral shRNA delivery. A and B, targeted post-transcriptional gene silencing of dicer was confirmed by Western blot (A) and real time PCR (B). C and D, Matrigel angiogenesis assay was used to evaluate the functional effects of dicer knockdown in vitro. Cells were stained with calcein-AM (C), and the area within the formed tubes was quantitated by analyzing three high powered fields per well using AxioVision Rel 4.8 software (D). Matrigel tube formation was compromised in dicer knockdown EOMA cells. E, reduced tumor size in response to dicer knockdown. F, incidence of tumor formation in mice injected with EOMA cells transfected with controls shRNA or dicer shRNA. G, tumor volume as quantified using calipers (length × width × height). The results are means ± S.D. of at least three independent experiments. *, p < 0.05.

Article Snippet: In Vitro Angiogenesis Assay Four-well plates were coated with 100 μl of Matrigel® (Cultrex® Basement membrane extract reduced growth factor; R&D Systems, Minneapolis, MN) and let to solidify for 30 min at 37 °C.

Techniques: Generated, shRNA, Western Blot, Real-time Polymerase Chain Reaction, Angiogenesis Assay, Functional Assay, In Vitro, Staining, Software, Injection, Transfection