Patient 3′UTR is associated with a significant reduction in endogenous FMRP and reporter activity in multiple cell types. (A) Photograph of a patient harboring the FMR1 c.*746C allele. (B) (Top) Representative Western blot of FMRP and eIF4e from two unrelated control male lymphoblastoid cell lines and the patient-derived lymphoblastoid cell line. (Bottom) The band density on the Western blot of three independent protein preparations was digitally quantified by ImageJ software (NIH). Data shown are the mean ± SD. (C) Luciferase assay in HEK293 and Neuro2a cell lines using vectors with a control 3′UTR or the patient 3′UTR. Each of three independent experiments was normalized to cotransfected Renilla luciferase activity; data shown are the mean ± SD. (D) Luciferase assay results of three independent experiments in HEK293 cells using vectors of the control, patient, and the patient’s half-brother. Data shown are the mean ± SD. Unpaired two-tailed t test, *P < 0.05; a.u., arbitrary units; n.s., not significant.