Figure 3.
Farnesylated lamin A proteins do not induce gross activation of p53 in AG13353 cells. (A) Quantitative RT-PCR was used to measure mRNA abundance for the p53 target genes P21, GADD45B, and IGFPBP3. mRNA inputs were normalized to the combination of three housekeeping genes (see Materials and Methods). For each gene the expression relative to vector controls is indicated. Error bars represent SD of six independent experiments. For each gene the p value of each sample compared with the vector control was ≥0.15 except for IGFBP3 where the p value for wild-type lamin A was 0.01 and the p value for L647R lamin A was 0.09 compared with vector controls. (B) Western analysis of p21 protein levels. Lysates of asynchronous, subconfluent cells were probed simultaneously for p21 and actin. To induce p21 protein levels, cells were treated with 5 μM camptothecin for 24 h.