IL-17A regulates autophagy and promotes osteoclast differentiation through the ERK/mTOR/Beclin1 pathway
Fig 5
Inhibition of autophagy attenuates IL-17A-mediated osteoclastogenesis.
A specific siRNA targeting Beclin1 mRNA was used to knockdown Beclin1 expression. (A) At 24 h post-transfection, the siRNA transfection efficiency was detected by western blotting. Protein expression was normalized against β-Actin. RAW264.7 cells and BMMs were treated with Beclin1-si or 3-MA in the presence of IL-17A (1 ng/ml) and RANKL (50 ng/ml) for 5 d (B, C) and following treatment, TRAP-positive multinucleated cells were quantified (D, E). At least four wells were used for each tested reagent. Cells treated without RANKL and IL-17A (CTR-N) were used as a negative control. Cells treated with RANKL and IL-17A (CTR-R) were used as a positive control and the values are expressed as the mean ± SD. *p < 0.05 compared with CTR-R. Scale bar, 100 μm.