Figure 1. RT² Profiler™ PCR Array screening and TRAIL expression in preosteoclast cells.

RAW 264.7 cells were cultured as ground based control (Xg) or under μXg for 24 h. Total RNA was isolated from these cells were screened for RT Profiler array of 84 cytokine/growth factors mRNA expression using real-time PCR. The Array data were normalized with housekeeping gene panel B2M, HPRT1, RPL13A, GAPDH & ACTB and differential gene expression represented as (a) Scatter plot analysis (Red—upregulated; Black—unchanged; Green—downregulated); (b) bar graph showing specific gene expression changes. (c) Mouse bone marrow derived non-adherent cells were subjected to Xg or μXg for 24 h. Total RNA isolated from these groups was further analyzed for TRAIL mRNA expression using real-time RT-PCR. The 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 Xg. (d) Confocal microscopy analysis for TRAIL expression in Xg or μXg subjected preosteoclast cells. Nuclear staining was performed with DRAQ5 and TRAIL expression was visualized by IX81 confocal microscope. The boxed areas are shown digitally zoomed at higher magnification (63×) in the lower panel.