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. Author manuscript; available in PMC: 2008 Feb 28.
Published in final edited form as: J Biol Chem. 2003 Aug 12;278(43):41862–41870. doi: 10.1074/jbc.M308336200

Fig. 5. Antisense Slug mRNA treatment restored the gene expression of Col2a1.

Fig. 5

A. The effect of antisense Slug mRNA was tested using HA-tagged Snail expression vector. Undifferentiated ATDC5 cells were transfected with 0.25 μg of HA-Snail expression vector together with antisense Slug mRNA vector. The dose of antisense Slug mRNA expression vector was 0 μg (lane 1), 0.125 μg (lane 2), and 0.25 μg (lane 3). The total amount of plasmids was adjusted to 2μg with pcDNA3.1 empty plasmid. The experiment was triplicated and a representative result is shown. B. Cotransfection of the reporter and antisense-Slug vectors with HA-tagged Snail expression vector recovered the suppressive effect in a dose-dependent manner. Undifferentiated ATDC5 cells were transfected with 977-luc vector, antisense Slug vector, and HA-tagged Snail vector. Then, 0.25 μg of antisense Slug mRNA vector was transfected. The dose of HA-Snail vector was 0 μg (lane 1 and 2), 0.05 μg (lane 3), 0.1 μg (lane 4), and 0.2 μg (lane 5). At the same time, the amount of HA-Snail Hg (lane 3), 0.1 μg (lane 4), and 0.2 μg (lane 5). At the same time, the amount of HA-Snail protein was checked by Western blotting. Protein was extracted 2 days after transfection from the cells in duplicated wells in the plate for luciferase assay.