All individuals carrying RAD50 LOF variants (Table 2) and deleterious missense variants with CADD score ≥ 25 present in familial ESCC including c.G2177A:p.R726H, c.C2287T:p.R763C, c.G3716A:p.R1239Q, and c.C3790T:p.L1264F (Table S1) were validated by Sanger sequencing, as shown in Figure 2 for representative validation of LOF variants c.C2014T:p.Q672X, and c.2165_2166insT:p.K722fs located at exon 13 containing the zinc hook domain and the missense c.C3790T:p.L1264F variant at exon 25. The gene discussed is RAD50; the disease is esophageal squamous cell carcinoma.