Western blot and immunohistochemical localization of ClCa to the olfactory cilia.

<p>A. Western blots of an enriched ciliary membrane preparation (OC), olfactory epithelium membranes (OE) and brain (positive control) samples using the anti-ClCa and the anti-actin antibodies. B. Immunolabeling of the olfactory epithelium (left panels) and of dissociated olfactory neurons (right panels) using specific anti-ClCa (green) and anti-CNGA2 (red) antibodies. Please note that in the dissociated cells the antibody immunoreactivity was found in the cilia, but there was also immunoreactivity in the cell body likely due to leakage of the ciliary proteins to the cell body after dissociation. Calibration bars in B: left, 10 µm; right top, center, 10 µm; right bottom, 2 µm. C and D. Immunolabeling of the olfactory and respiratory epithelia by anti-ClCa, anti-ANO2 known to label the olfactory cilia <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0069295#pone.0069295-Billig1" target="_blank">[16]</a> and anti-acetylated tubulin that labels the cilia layer in both the olfactory and respiratory epithelia. Bars are 50 µm. C. The ciliary layer of the olfactory epithelium is labeled by both anti-ClCa (green) and anti-ANO2 (red). D. Both the respiratory and olfactory ciliary layers are labeled by anti-acetylated tubulin (green). The olfactory epithelium is identified by labeling with anti-ANO2 (red).