Figure 2

Suppressive effect of ZSTK474 on OC precursor cells. a) Inhibition of Akt phosphorylation by ZSTK474. RAW264.7 cells were incubated with or without 0.1 μM of ZSTK474 for 30 minutes and for another 15 minutes in the presence of soluble RANKL. The phosphorylated Alt (p-Akt) and whole Akt (Akt) were examined by the Western blotting. b) The expression of NFATc1 was determined in RAW264.7 cells cultured for 48 hours in the presence of soluble RANKL with or without ZSTK474 pretreatment. c) NFATc1 localization was visualized using immunofluorescence microscopy in RAW264.7 cells cultured with sRANKL and TNF-α for 24 hours. Treated with vehicle (left) and 0.1 μM of ZSTK474 (right). d) The phosphorylation of Akt in RAW264.7 cells cultured with soluble RANKL and TNF-α was inhibited by ZSTK474. e) RAW264.7 cells were cultured in the presence of RANKL and TNF-α in the presence or absence of ZSTK474. The number of TRAP staining-positive multinucleated cells was counted.