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. 2015 May 11;10(5):e0125993. doi: 10.1371/journal.pone.0125993

Fig 5. Validation of the BbTPx-1 expression in the transfectants.

Fig 5

Expression of Bbtpx-1 in the wild type parental strain (WT), Bbtpx-1 KO parasites (KO), Bbtpx-1 genome integrant (GI), and Bbtpx-1 episomal transfectant (E). (A) RT-PCR assays were performed for Bbtpx-1 gene, methionyl-tRNA synthetase gene (BbMetRS), and a negative control without reverse transcription (RT—(Bbtpx-1)). M, 100 bp DNA ladder marker. Expected RT-PCR band size of 133 bp and 157 bp are indicated for Bbtpx-1 and BbMetRS, respectively. (B) Indirect immunofluorescence assay with anti-BbTPx-1 serum. Differential interference contrast and fluorescence images are merged. The red fluorescence in the cytoplasm of the complemented transfectants indicates expression of BbTPx-1. (C) Western blot analysis with anti-BbTPx-1 serum. A band was detected from WT, GI, and E, which is consistent to the expected size of BbTPx-1 protein of 22 kDa. The CBB staining image in the left panel shows equal loading.