Figure 6

Salmon hnRNPA0 binds CpG-B ODNs and colocalizes with CpG-B ODNs. A, Western blot with mouse anti-hnRNPA0 antibody confirms that the salmon homolog expressed in mononuclear phagocytes interacts with CpG-B ODNs. The experiment was performed as in Figure 1A. B, Transgenic salmon hnRNPA0 is detected both in cell nuclei and cytoplasm and it colocalizes with fluorescent CpG-B. CHSE cells were transfected with a plasmid expressing an EGFP-hnRNPA0 fusion protein (green) and stimulated with Cy5-CpG-B as in Figure 2. The transgenic hnRNPA0 was observed mostly in cell nuclei (arrowhead); however, in cells which appear to be in a process of cytokinesis (boxed area), hnRNPA0 can be observed in cytoplasmic granules where it colocalizes with Cy5-CpGB (Arrow). The cell nuclei were counter-stained with 7-AAD (blue pseudocolor). Note that the emission, but not the excitation, spectrum of 7-AAD overlaps with that of Cy-5 and therefore, this channel detects both the genomic DNA and the endocytosed CpGs.