Effects of intermittent hypoxia exposure to neuroblastoma cells on CaSR expression in RAW 264.7 cells.

<p>(<b>A</b>) RAW 264.7 cells were exposed to growth medium (control), growth medium with RANKL or CM from parental SH-SY5Y cells (N), or intermittent hypoxia-exposed (IH) cells and cultured for six days under normoxia. The medium was changed after three days. Cells were fixed and labeled with CaSR antibodies and Alexa-594 antimouse-conjugated antibodies. Nuclei were stained with DAPI. RANKL-treated RAW cells served as a positive control for CaSR expression during osteoclastogenesis. A representative image is shown. Bar: 100 µm. (<b>B</b>) Cell lysates were analyzed for the levels of CaSR protein by Western blotting. The band intensity was measured and the protein level was normalized to the corresponding β-actin level. The results are expressed as relative quantity to the control (first lane of the blot).