In each panel, the distribution of fitness effects (DFE) of all possible loss-of-function (LOF) mutations is denoted with a black curve. For n DNMs in a disease cohort, the gray lines denote 100 bootstrapped DFEs of a set of n DNMs randomly sampled from the full set of LOF mutational opportunities. The estimated DFE of de novo LOF mutations in (A) affected males from the Deciphering Developmental Disorders (DDD) cohort, (B) affected females from the DDD cohort, (C) affected males from the Simons Simplex cohort, (D) affected females from the Simons Simplex cohort, (E) affected males from the MSSNG cohort, excluding multiplex families, (F) affected females from the MSSNG cohort, excluding multiplex families, (G) affected males from the SPARK cohort, (H) affected females from the SPARK cohort, (I) affected males in multiplex families from the MSSNG cohort, and (J) affected females in multiplex families from the MSSNG cohort.