Fig. 4

Functional analysis of the COL4A2 variant effect on mRNA splicing. (a) Construction strategy of pcDNA3.1 minigene vector, * representative of the variant site. (b) The sequencing of wildtype COL4A2 gene vector and c.549 + 5G > A COL4A2 variant gene vector. (c) Gel electrophoresis of reverse transcription polymerase chain reaction products displayed a single band a (estimated 410 bp) from the wild type (wt) and a smaller band b (estimated 338 bp) in the mutant type (mut). (d) Illustration of the sequencing of band a (wild type in HEK293T and Hela cells) and band b (variant c.549 + 5G > A in HEK293T and Hela cells) products lead to a shorter transcript with deletion of exon 8 including 72 bp