Figure 4.
GGT Activity and LH3 Protein Level of the Proband
GGT activity in proband's serum (A) and in lymphoblastoid cells (B). The activity in the cells is expressed per μg soluble protein and compared with a mean value of two control cell lines. The activity in serum is expressed per μl serum and compared with a mean value of five control sera. The SD is indicated by the bars. LH3 protein level was analyzed by western-blot analysis (C) from lymphoblastoid cells of the patient (P) and from three control lymphoblastoid cells (C1-C3). A total of 2 mg soluble protein from lymphoblastoid cells was pulled down with Concanavalin A-sepharose (GE Healthcare) and applied to the lanes. Molecular-weight markers are indicated. A GGT activity assay was performed as described elsewhere.28 The proteins were fractionated under reducing conditions by 7.5% SDS-PAGE, transferred onto an Immobilon-P membrane (Millipore), and incubated with a polyclonal PLOD3 antibody (Proteintech Group). Anti-rabbit-IgG-HRP (P.A.R.I.S biotech) was used for secondary detection with ECL+ system (GE Healthcare) and Biomax XAR X-ray film (Kodak).