Figure 1

Western blots of patient fibroblasts with antibodies against LIPT1 (A), lipoic acid (B), and tubulin (Sigma-Aldrich) or actine in cultures in basal conditions (A, B) and after LIPT1 transfection (C). A and B: Control, patients (P) with PDHA1, NFU1 and LIPT1 mutations were as indicated. A: the LIPT1 antibody failed to detect the LIPT1 protein in patient fibroblasts. B: Anti-lipoate antibody failed to detect the expected lipoylated proteins of ketoacid dehydrogenases (PDHc, α-KGDHc and BCKDHc) in the patient fibroblasts as well as in fibroblasts of another patient with NFU1 mutations, whereas normal bands were seen in patient fibroblasts with PDHA1 mutations and in control. C: Control (C), Patient with LIPT1 mutations (P), patient fibroblasts transfected with LIPT1 (P + LIPT1). Anti-lipoate antibody revealed normalized and very moderately increased amounts of BCKDH (x8) and α-KGDH (x1.8) proteins, respectively, in patient fibroblasts after LIPT1 transfection whereas no band was detected for PDH protein.