Fig. 4

Splicing pattern evaluation by in vitro minigene assays. a Non-truncating mutations in FLCN were divided into four groups for splicing evaluation. None of these non-truncating mutations led to an abnormal transcript, compared with those of the wild-type minigenes. While, the positive control, variant c.249 + 1G > A, yielded a shorter transcript as expected. b Sanger sequencing of the aberrant transcript produced by c.249 + 1G > A showed that this mutation caused a partial deletion of 125 bp in FLCN exon 4, consistent with the in vivo results from patient 9–1