Fig. 4

EVC2 variants caused EVC in case 49. A The image of US examination of case 49 revealed short and curved long bones. B The pedigree map of case 49. C MLPA results of EVC2 in case 49: a heterozygous deletion of EVC2-22 ~ EVC-8. D–F Sanger sequencing showed that the proband had a novel variant inherited from the mother. G Schematic presentation of the gross deletion. Arrows indicate genes; Solid horizontal lines indicate retained regions; Broken horizontal lines indicate deleted regions; Multiple primers were designed to be regularly spaced every ~ 25 kb (step 1), ~ 5 kb (step 2), and ~ 1.5 kb (step 3) within breakpoint regions. Long-range PCR with the forward primer of the D131 and the reverse primer of the U4 generates a PCR product of ~ 1.5 kb. H The breakpoint sequences of case 49 and Sanger-sequence of the gross deletion. I UCSC Genome Browser information around the telomeric breakpoint regions (chr4: 5,547,999–5,548,399)