Skip to main content
. 2008 May 21;3(5):e2236. doi: 10.1371/journal.pone.0002236

Figure 4. Treatment of MYCN-amplified LAN-5 cells with antagomir-17-5p inhibits in vitro tumorigenesis through p21 and BIM upmodulation.

Figure 4

(A) Colony formation of LAN-5 cells treated with antagomir-17-5p or a control antagomir. 24 h after treatment, cells were plated in a soft agar semisolid medium and colonies were counted after 2 weeks. In each experiment, cells were plated in triplicate. Mean±s.d. (n = 3). (B) Cell cycle analysis of LAN-5 cells treated with antagomir-17-5p or a control antagomir. After treatment, cells were starved for 24 h and then incubated with a complete medium for 16 h before BrdU incorporation and FACS analysis. Percentage of cells in G1, S or G2-M phase of the cell cycle is indicated. A representative experiment is shown. (C) Western blot (left panel) and qRT-PCR (right panel) of p21 expression in LAN-5 cells treated with antagomir-17-5p or a control antagomir. A representative Western blot is shown. Mean±s.d. (n = 3). (D) Apoptosis of LAN-5 cells treated with antagomir-17-5p or a control antagomir. 24 h after treatment, cells were incubated with Annexin V and Cytox Green and analyzed by FACS. A representative experiment is shown. (E) Western blot (left panel) and qRT-PCR (right panel) of BIM expression in LAN-5 cells treated with antagomir-17-5p or a control antagomir. A representative Western blot is shown. Mean±s.d. (n = 3). (F) Luciferase activity in Tet-21/N cells transfected with pGL3-prom-BIMUTR wt or mut in combination with a control or an anti-miRNA oligonucleotide complementary to miR-17-5p. The ratio of normalized luciferase activity in pGL3-prom-BIMUTR wt versus mut transfected cells is indicated. Mean±s.d. (n = 6). * P<0.05, ** P<0.01, *** P<0.001.