A-type lamins in primary fibroblasts from HGPS and unaffected controls. (a) Western blot experiments with antibodies to lamins A (left panel), C (middle panel), and A/C (right panel). (b) RT-PCR screening with exon (ex) 7/8 or ex9/11, and ex12 primer showed fragments corresponding to lamins A (640 base pairs, bp) and AΔ150 (490 bp), or lamins AΔ10 (319 bp) and AΔ10, Δ150 (169 bp), respectively. RT-PCR with primers in ex1, 2, or 3, and ex10, resulted in fragments corresponding to lamin C (1612, 1350, and 1118 bp, respectively). Asterisk (*) indicates that the primer was located in the region of exon 10 specific for lamin C. Negative control lanes contained PCR samples with genomic DNA, no RT, and no template controls (M, 100 bp DNA ladder (Invitrogen)). (c) Representative western blot with protein extracts from multiple passages of fibroblast cell cultures. (d) Changes in protein levels during passaging illustrated as the ratio of normalized band intensities for late and early passage protein extracts of the same culture. Results represent means ±SD.