Fig. 5.

Inhibitory effect of miR-26a is rescued by CTGF. (A–D) BMMs were cultured for 2 days in the presence of M-CSF and RANKL. Two day-old pre-OCs were transfected with miR-26a mimic, miR-26a inhibitor, or negative control as indicated. Transfected cells were further cultured with M-CSF and RANKL in the presence or absence of CTGF (10 ng/ml). (A) Cultured cells were fixed and stained for TRAP. (B) Numbers of TRAP + MNCs were counted. (C) Cultured cells were fixed and stained with phalloidin. (D) Numbers of MNCs containing an actin ring were counted. (E, F) BMMs were cultured on Osteo Assay plates for 2 days in the presence of M-CSF and RANKL. Two-day-old pre-OCs were transfected with the miR-26a mimic, miR-26a inhibitor, or negative control as indicated. Transfected cells were cultured for 36 h with M-CSF and RANKL in the presence or absence of CTGF (10 ng/ml). (E) Resorption lacunae were visualized by bright-field microscopy. (F) Numbers of resorption pits were counted. Data represent means ± SD of triplicate samples. **p < 0.05, ***p < 0.001 vs. control. Results are representative of at least three independent sets of similar experiments