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. 2015 Apr 7;10(4):e0122256. doi: 10.1371/journal.pone.0122256

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© 2015 Chen et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Fig 2 — (A) Southern blot analysis of Ha-annexin. Genomic DNA from H. avenae was digested with BamHI and EcoRI, respectively, and probed with the Dig-labeled CDS of Ha-annexin. They had 3 and 2 signal strips, respectively. (B) In situ hybridization of the Ha-annexin transcripts in pre-parasitic second-stage juveniles. Signal of antisense Ha-annexin DIG-Labeled cDNA probes localized within the subventral glands (SVGs), with sense probes as a negative control. The SVG, metacorpus (M), and stylet (S) are indicated with arrows. Scale bar = 20 μm. (C) Developmental expression pattern of Ha-annexin. The relative expression level of Ha-annexin was quantified using qPCR for six different H. avenae stages. The fold change values were calculated using the 2^-ΔΔCt method and presented as the change in mRNA level in various nematode developmental stages relative to that of egg. Each column represents the mean of 3 independent assays with standard deviation. preJ2: pre-parasitic second-stage juvenile; parJ2, J3 and J4: parasitic second-, third- and fourth-stage juvenile, respectively.