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. 2022 Feb 18;60(1):467–478. doi: 10.1080/13880209.2022.2039216

Figure 2.

Figure 2.

Effect of FAEE on RANKL-stimulated osteoclast formation of F-actin rings in osteoclasts and bone resorption. In the assay to measure formation of F-actin rings, RAW 264.7 cells were plated into 12-well plates and incubated with FAEE (25, 50, or 100 μM) in the presence of RANKL for 5 days. Cells were fixed and stained with fluorescein rhodamine phalloidin to observe F-actin rings (magnification: 200×) and were merged by Image J software (A), the number of F-actin rings were calculated under a fluorescence microscope (C). In the pit-formation assay, RAW 264.7 cells were seeded on an Osteo Assay Surface multi well plates. Surfaces were treated similarly and cultured for 5 days. Resorption area was captured using an inverted microscope (magnification: 100×) (B). Pit areas were quantified by Image J software (D). C-terminal telopeptide of type I collagen (CTX-I) concentration in cell supernatants using Enzyme-linked immune-sorbent assay (ELISA) (E). Data are the mean ± SEM of triplicate experiments. ## p < 0.01 and ### p < 0.001 versus RAW 264.7 cells without RANKL, ** p < 0.01 and *** p < 0.001 versus RAW 264.7 cells with RANKL alone.