Research Paper Volume 15, Issue 19 pp 10732—10745

Figure 5. Micheliolide inhibited RANKL-induced activation of the p38 MAPK signaling pathway during osteoclast differentiation. BMMs were plated at the same density of 2.5*10³ cell/cm in 4 different wells. 2 wells received DMSO treatment (Con-1, Con-2) and another 2 wells received 20 uM Micheliolide (MCL-1, MCL-2) treatment in the presence of M-CSF and RANKL for 5 days. Then, samples were collected and sent for RNA-seq. (A) Heatmap of the total 23947 detected genes in control and Micheliolide treatment group. (B) Volcano Plot of the detected genes. (C) Heatmap of osteoclast related genes detected in control and Micheliolide treatment group. (D) GO analysis of the RNA-sequencing. (E) BMMs were pre-treated with DMSO or Micheliolide (20 uM) for 2 hours, then stimulated with RANKL for indicated times (0, 5, 15, 30, 60 min). Proteins were collected and WB for MAPK signaling pathway was performed. All data: mean ± SD, n=5, *p < 0.05 comparing to RANKL+DMSO group, **p < 0.01 comparing to RANKL+DMSO group, ***p < 0.001 comparing to RANKL+DMSO group, N.S. no significant difference comparing to RANKL+DMSO group.