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. 2019 Apr 22;47(11):5922–5935. doi: 10.1093/nar/gkz259

Figure 4.

Figure 4.

Increased usage of the endogenous alternative splice site in CRL 1474 cells following ASO treatment. (A) Diagram indicating the target location of ASO1919 on the LMNA WT structure model, highlighted by a green line. The alternative 5′ SS sequence is highlighted by a blue line. (B) Diagram indicating the target location of ASO074 (red) and ASO1919 (green) in the context of the splicing cassette. The locations of the forward and reverse primers used to measure endogenous expression of LMNA/Δ150 are indicated. (C) RT-PCR analysis for LMNA/Δ150 expression levels of each treatment group. CRL 1474 cells were transfected with 25–100nM of ASO1919, 25nM of ASO074, or 25nM of scrambled ASOSCR control. RNA was harvested at 72 hours post transfection. TBP expression served as a loading control. (D) Quantitative PCR results for LMNA and Δ150 expression levels for the same samples tested in panel C. Expression levels are normalized to expression of TBP. Data are expressed as mean ± SEM. N.S: nonspecific PCR product. ASO: antisense oligonucleotide; UT: untransfected; SCR: scrambled ASO; NC: negative control (PCR reaction mix without cDNA template).