Fig. 2

Antisense oligonucleotide (AON)-mediated rescue for intronic ABCA4 splice variants. For three deep-intronic ABCA4 variants (c.859–540C>G, c.5197–557G>T, and c.4539+1106C>T), AON rescue experiments on transfected HEK 293-T cells were undertaken (a–c). AON experiments were also performed on the fibroblasts of a patient with c.[4539+1106C>T];[6089G>A] and control fibroblasts (d). A gel image is shown on the left, depicting the reverse transcription polymerase chain reaction (RT-PCR) product derived from mutant (MT) and wild-type (WT) midigene construct after transfection with three different AONs (HEK: untransfected HEK 293-T, MQ: negative control PCR, NT: nontransfected cells, SON: sense oligonucleotide). Rhodopsin (RHO) and actin (ACTB) were used as loading controls for RT-PCR products derived from experiments on HEK 293-T cells and fibroblasts, respectively. In fibroblasts, cycloheximide (CHX) was added. On the right, resulting RT-PCR products after AON rescue experiments are semiquantified using capillary analysis, using the ratio WT transcript/transcript including pseudo-exon (PE). If multiple PE containing transcripts were formed, they are grouped together in one PE group.