Mutation screening
We obtained DNA samples for each patient from peripheral blood lymphocytes by standard procedures (Sambrook, Russell, & Fritsch, 2001). All 11 coding exons and intron-exon boundaries of EFHC1were amplified by polymerase chain reaction (PCR; primer sequences available upon request). Sanger sequencing was performed by capillary electrophoresis in an ABI 3500xL Genetic Analyzer using the BigDye® Terminator Cycle Sequencing Kit (Thermo Fisher Scientific, Waltham, MA, USA). Sequence variants were described according to the conventional nomenclature (den Dunnen & Antonarakis, 2001; den Dunnen, 2019) based on the full-length EFHC1 isoform (NM_018100) and deposited in a public genomic database of GGEs (http://bipmed.iqm.unicamp.br/GGE).