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Figure 3 | BMC Molecular Biology

Figure 3

From: Natural antisense RNA inhibits the expression of BCMA, a tumour necrosis factor receptor homologue.

Figure 3

Antisense BCMA RNA affects BCMA protein expression only on translational level. 293T cells were transfected with insert-less vector or vector expressing FhBCMA in the presence of various amounts of p12-ATG expression vector. 48 hours after transfection cells were harvested and the cell suspension split in two. One half (about 107 cells) was lysed in appropriate buffer and 14 μg of the resulting cell lysate was assayed by western blotting using the M2 anti-FLAG monoclonal antibody to test for FhBCMA. We tested that the protein levels were similar in the various samples by western blotting using a rabbit polyclonal anti-actin antibody. The other half of the cell suspension was used to prepare total RNA, which was used for northern blot to assess BCMA and human β-actin levels. 293T cells were transfected with (A) none, (B) 1 μg of insert-less pcDNA3, (C) 100 ng hBCMA, (D) 100 ng p12-ATG, (E) 100 ng hBCMA and 100 ng p12-ATG, (F) 100 ng hBCMA and 900 ng p12-ATG. Except for (A), cells were transfected with total of 1 μg of plasmid using when needed insert-less pcDNA3.

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