CENP-A overexpression and knockdown in HepG2 cells.

Analysis of CENP-A mRNA (A, C) and protein (B, D) levels in HepG2 cells transfected with CENP-A-expressing plasmids or siRNA-expressing plasmids, as described in Materials and methods. A: RT-PCR (left) and qPCR (right) measurement of CENP-A transcript in cells transfected with pcDNA3.0-CENP-A or pcDNA3.0 empty vector. **Pt-test; n = 3. B, left: Representative Western blots for CENP-A in cells treated as in A. Right: Quantification of the blotting shown in the left panel. **Pt-test; n = 3. C: RT-PCR (left) and qPCR (right) examination of CENP-A mRNA levels in HepG2 cells transfected with CENP-A1, CENP-A2, or pSil2.1-U6 empty vector (control). **Pn = 3. D: Immunoblot of cells treated as in C. Left: Representative Western blots for CENP-A. Right: Quantification of the blotting shown in the left panel. **Pn = 3. GAPDH was used as an internal control in all the experiments.