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. 2016 Mar 15;6:22389. doi: 10.1038/srep22389

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Copyright © 2016, Macmillan Publishers Limited

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(A) BMMs were pretreated with Ewha-18278 for 1 hour, the cells were stimulated with RANKL for the indicated times. The cell lysates were subjected to immunoblot analysis. RANKL signaling pathways assessed by phosphorylation of ERK, p38, JNK, and IκBα. Immunoblots were stripped and then reprobed with total ERK, p38, JNK, and IκBα. (B,C) BMMs were treated with Ewha-18278 in the presence of M-CSF and RANKL for the indicated periods. Total RNA was extracted from the cells and used in real-time PCR to quantify the mRNA levels of NFATc1 (B) and Atp6v0d2 (C) genes. The relative expression levels were normalized to the level of actin mRNA as an internal control gene. *P < 0.05. Data represent mean ± SD. (D) NFATc1 and Atp6v0d2 expression was determined by immunoblot analysis. Actin serves as a loading control.