Figure 4. mRNA and protein expression of αTFP in control and/or BTHS lymphoblasts transfected with αTFP, MLCL AT activity in control and BTHS lymphoblasts transfected with αTFP, MLCL levels in BTHS lymphoblasts transfected with αTFP RNAi and mitochondrial respiratory chain subunits in BTHS lymphoblasts transfected with αTFP.

Age-matched control lymphoblasts (A) or BTHS lymphoblasts (B) were mock transfected (Mock) or transfected with αTFP plasmid (αTFP) and mRNA expression of αTFP determined as described in Materials and Methods. Data represents the mean ± standard deviation of four experiments. *p<0.05. C. BTHS lymphoblasts were mock transfected (Mock) or transfected with human recombinant αTFP plasmid (αTFP) and mitochondrial fractions prepared and 150 µg protein subjected to SDS-PAGE followed by western blot analysis using the anti-αTFP antibody as described in Materials and Methods. A representative blot is depicted. D. Control (Mock) and BTHS (BTHS) lymphoblasts were mock transfected or transfected with αTFP plasmid (+αTFP) and MLCL AT activity determined. Data represent the mean ± standard deviation of three experiments. *p<0.05. E. BTHS lymphoblasts were mock transfected (Mock) with off-target αTFP nucleotide sequence or transfected with two different on-target αTFP RNAi nucleotide sequences (RNAi1, RNAi2) and CL and MLCL levels determined. Data represents the mean of two experiments. F. BTHS lymphoblasts were mock transfected (Mock) or transfected with human recombinant αTFP (αTFP) and mitochondrial fractions prepared and 40 µg protein subjected to SDS-PAGE followed by western blot analysis for respiratory chain protein subunits as described in Materials and Methods.