Figure 5.

Interleukin (IL)-7 induced osteoclast formation through a signal transducer and activator of transcription 5 (STAT5) signaling pathway. (A) Control (RAW-vector) and RAW-IL7ROE cells were stimulated for the indicated time intervals with IL-7 (10 ng/mL). Then, immunoblot analysis was performed. Results are representative of three independent experiments. (B) RAW-IL7ROE cells were stimulated for 30 min with RANKL (50 ng/mL) and/or IL-7 (10 ng/mL). Then, immunoblot analysis was performed. Results are representative of two independent experiments. (C) RAW-vector or RAW-IL7ROE cells were transfected with wild-type STAT5a (STAT5A-WT), constitutively active STAT5a (STAT5A-CA), and dominant-negative STAT5a/b (STAT5A-DN, STAT5B-DN), then cultured for 6 days with RANKL (50 ng/mL) and/or IL-7 (2 ng/mL), replacing the medium at 3-day intervals with fresh cytokines. TRAP staining was performed as described in Figure 1. Representative images showing TRAP⁺ cells on day 6 are shown. Results are representative of two independent experiments. (D) Cells were cultured on top of dentine disks in 96-well culture plates in the above conditions for 14 days. Then, surface roughness was analyzed as described in Figure 1. Representative confocal images are shown. The arrowhead indicates the resorption pit. Results are representative of two independent experiments.