Research Paper Volume 13, Issue 20 pp 23652—23671

Up-regulation of SIRT1 induced by 17beta-estradiol promotes autophagy and inhibits apoptosis in osteoblasts

Detection of caspase-3 and Bcl-2 expression in hFOB1.19 osteoblasts by real-time PCR and Western blot assay. Cells were pretreated with or without SRT1720 and EX527 for 2 h, and then cultured with 17β-E2 (10-6 M) for 24 h. The expression of caspase-3 (A) and Bcl-2 (B) mRNA in osteoblasts were assessed by RT-PCR. The data were normalized to GAPDH. The expression level of caspase-3 (C) and Bcl-2 (D) proteins in osteoblasts were measured by Western blotting. The histograms showed the relative protein expression of caspase-3 (C) and Bcl-2 (D) normalized to β-actin protein, and the densitometry analysis of the blots were estimated by using the Image J Software. The data were expressed as the means ± SD from 3 independent experiments. Statistically significance was evaluated using the Student’s t-test. * P  0.05 vs. the control group was considered significant.

Figure 7. Detection of caspase-3 and Bcl-2 expression in hFOB1.19 osteoblasts by real-time PCR and Western blot assay. Cells were pretreated with or without SRT1720 and EX527 for 2 h, and then cultured with 17β-E2 (10-6 M) for 24 h. The expression of caspase-3 (A) and Bcl-2 (B) mRNA in osteoblasts were assessed by RT-PCR. The data were normalized to GAPDH. The expression level of caspase-3 (C) and Bcl-2 (D) proteins in osteoblasts were measured by Western blotting. The histograms showed the relative protein expression of caspase-3 (C) and Bcl-2 (D) normalized to β-actin protein, and the densitometry analysis of the blots were estimated by using the Image J Software. The data were expressed as the means ± SD from 3 independent experiments. Statistically significance was evaluated using the Student’s t-test. * P < 0.05 vs. the control group was considered significant.