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. 2020 Aug 4;42(5):744–754. doi: 10.1038/s41401-020-0477-4

Fig. 5. ORI suppressed the RANKL-induced PLCγ-NFATc1 signaling pathway in RAW264.7 cells.

Fig. 5

a RAW264.7 cells stably expressing the NFAT-luc luciferase reporter construct were pretreated with ORI and CsA and then stimulated for 24 h with RANKL (100 ng/mL). Then, the luciferase activity was measured. b RAW264.7 cells were cultured as described above, and real-time PCR was performed on cells stimulated with or without RANKL and ORI for 24 h to examine NFATc1 mRNA expression. c–f Total and nuclear extracts from RAW264.7 cells pretreated with ORI and stimulated with RANKL (100 ng/mL) for 24 h were analyzed by Western blotting with antibodies against NFATc1, lamin A/C, and β-actin. g, h RAW264.7 cells were pretreated for 30 min with ORI, stimulated with RANKL for 30 min, and analyzed by Western blotting with p-PLCγ1 (Tyr 783), β-actin, and PLCγ1 (1249) antibodies. ##P < 0.01, ###P < 0.001 vs the control group; *P < 0.05, **P < 0.01, ***P < 0.001 vs the RANKL-treated group.