Fig. 1 |. Design and testing of a PPMO-based strategy for blocking the production of progerin.

a, Right, Normal LMNA transcript processing. Left: aberrant LMNA splicing. The HGPS G608G mutation position is indicated by the arrow at the junction of the retained exon 11 sequence (dark gray) and the 150-nt portion of exon 11 that is deleted (light gray) by splicing at the cryptic splice site. The antisense PPMO binds transcripts containing the mutation, thereby blocking cryptic splice site recognition by the spliceosome. b, Tiling of 11 alternatively designed PPMOs around the cryptic splice site generated by the LMNA G608G (c.1824C>T) mutation. The two controls included an oligonucleotide complementary to the normal LMNA sequence (wtLMNA, WT) and an oligo predicted to lack complementarity to any sequence in the human genome (scrambled control, CTRL). c, RT–qPCR results, normalized relative to TFRC RNA, of proband-derived fibroblasts cultured in the presence of 6 μM of alternative PPMOs for two weeks. Averages and s.d. were determined from triplicate reactions of biological triplicates. d, Immunoblots of triplicate lysates from proband-derived fibroblasts cultured in the presence of 6 μM of alternative PPMOs for two weeks. e, Quantitation of lamin A, progerin and lamin C proteins from western blot analysis, normalized relative to β-actin (ACTB). Values and error bars represent the mean ± s.d. of triplicate cultures. A, lamin A; P, progerin; C, lamin C; *P < 0.05, **P < 0.01,***P < 0.001 versus transcripts or protein from cells receiving saline.