Figure 2

Generation of a dTDP-43 null allele. (A) Schematic representation of the truncated dTDP-43 protein caused by the single nonsense nucleotide mutation at amino acid 367. (B) Sequencing of the PCR fragment amplified from the genomic DNA of heterozygous flies confirmed the single nucleotide mutation from G to A. (C) Western blot analysis of protein lysates from the heads of wildtype (WT) or dTDP-43^{Q 367X}homozygous mutant adult flies confirmed the absence of full-length dTDP-43 in mutants. The asterisk indicates a nonspecific band recognized by this antibody. The rabbit polyclonal antibody was raised against the peptide fragment spanning amino acids 179-192 and did not recognize the putative truncated dTDP-43 in mutants, indicating instability of the mutant dTDP-43 mRNA or protein. (D) qRT-PCR analysis of dTDP-43 mRNA levels in wildtype and dTDP-43^{Q 367X}mutant flies. Two pairs of primers (Additional file 2) used for qRT-PCR yielded similar results. Values are mean ± SD. *P < 0.05.