Fig. 2: EXOC7 exon 7 splice acceptor variant disrupts splicing. | Genetics in Medicine

Fig. 2: EXOC7 exon 7 splice acceptor variant disrupts splicing.

From: Regulation of human cerebral cortical development by EXOC7 and EXOC8, components of the exocyst complex, and roles in neural progenitor cell proliferation and survival

Fig. 2

(a) Top, diagram of EXOC7 human minigene construct with splice acceptor variant. Blue arrows mark reverse transcription polymerase chain reaction (RT-PCR) primers used to generate complementary DNA (cDNA) products shown in gel image. Arrows from each band in gel point to splicing diagram determined by Sanger sequencing of the cDNA. Wild-type (WT) minigene generated two in-frame isoforms (left), whereas variant minigene generated one in-frame isoform and two novel out-of-frame isoforms (right). Two variant isoforms encode novel stop codons that lead to premature protein truncation. Asterisk (*) indicates low-abundance product that could not be subcloned for sequencing. (b) Immunoblot of EXOC7 protein in WT and variant HAP1 cells showing reduction of two EXOC7 isoforms. GAPDH is a protein loading control. (c) Quantification of (b) showed significant 50% reduction of larger EXOC7 band (band 1, p = 0.045), while the lower weight band (band 2) was not significantly reduced. P values calculated by two-tailed t test. Error bars represent SEM.

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