Fig. 3

Effect of the splicing mutation on ELAC2 expression and on selected mitochondrial genes. a) The amplification products of ELAC2 cDNA from patients, controls and parents were seen on a 2 % agarose gel. Bright bands were detected in the lanes of two healthy controls (C1 and C2) and the mother (IV4) at 500 bp (according to the DNA size marker M). Multiple fainter bands were seen in the patients’ (V10 and V11) lanes suggesting diminished expression of the normal WT transcript and the presence of other abnormal splicing products. b) Expression analysis of ELAC2 protein in patient fibroblasts. Total protein lysates from patient (V10) and two different control fibroblasts were analyzed for ELAC2 protein expression by immunoblotting against an antibody specific for ELAC2 isoform1. HEK293T cell lysate was used as a positive control. Mouse Tubulin antibody was used as a loading control. The levels of the protein was negligible in patient fibroblast compared to the control. Densitometric analysis of ELAC2 protein bands normalized to Tubulin levels, revealed that ELAC2 protein expression in patient fibroblasts was 14 % of that detected in control fibroblasts. c) A significant difference is seen between relative expressions of different unprocessed mitochondrial transcripts mtATP8, mtND2, and mtND4, in skin fibroblasts from patient V10, compared to four different control samples (Ctl1, Ctl2, Ctl5, and Ctl6). The mRNA expression values were normalized to an internal control HPRT. X axis depicts quantitative expression; Y axis represents bar chart for controls, and patient samples, respectively