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. Author manuscript; available in PMC: 2015 Apr 3.
Published in final edited form as: Bone. 2014 Jan 23;61:125–131. doi: 10.1016/j.bone.2014.01.004

Fig. 4.

Fig. 4

(A) Autophagy inhibitor 3-methyladenine (3-MA) suppresses Atg5 and LC3 expression. Mouse non-adherent bone marrow cells were cultured in μXg for 24 h and treated with or without RANKL (75 ng/ml) and 3-MA (2 mM) for 24 h. Total RNA isolated were subjected to real-time RT-PCR analysis using gene specific primers for (A) Atg5 and LC3. (B) Cathepsin K gene expression. mRNA expression was normalized with respect to β-actin amplification. Each bar represents the mean ± SD of three independent experiments. *Significant (P < 0.05) difference when compared to ground based control without RANKL treatment. (C & D) RANKL stimulation and 3-MA inhibition of CREB activation under Xg and μXg conditions. Mouse non-adherent bone marrow cells were cultured in μXg for 24 h and treated with RANKL (75 ng/ml) in the presence and absence of 3-MA (2 mM) for different time points (0–120 min). Total cell lysate obtained from these samples was subjected to western blot analysis for p-CREB expression. Total CREB expression served as control.