Figure 4. Minigene constructs and products obtained in the in vitro splicing assay. A: Shown is a schematic representation of the constructs, which include TULP1 exons 12 to 15 (represented by boxes), flanked by 79–183 bp of intronic sequences (represented by straight lines). Constructs
harbored either the wild-type (WT) or one of two mutant alleles at the donor splice-site of intron 14: c.1495+2_1495+3insT
and c.1495+1G>A. Also shown are the locations of primers used for RT–PCR analysis (indicated by arrows) and the obtained splicing
products. Sizes of exonic and intronic fragments are indicated below them. B: WT and mutant constructs were transfected into Y79 cells, followed by RNA extraction and RT–PCR analysis. No PCR products
were obtained from cells transfected with the empty pCMV-script vector. β-actin (ACTB), a 437 bp product, served as an internal control for RNA quality and quantity.