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. Author manuscript; available in PMC: 2014 May 1.

Published in final edited form as: J Bone Miner Res. 2013 May;28(5):1214–1228. doi: 10.1002/jbmr.1851

Fig. 8 — (A) Senescence-associated β-galactosidase staining of WT (+/+) and Ercc1^−/ΔBMSCs that were treated with DMSO (vehicle), or the inhibitor of NF-κB activation IKKiVII (100nM, or 300nM) for 3 days. The percent of positive cells was counted (Right panel, n=4). Scale bar, 100μm. (B) qRT-PCR analysis of expression of osteoblastic markers in WT (+/+) and Ercc1^−/ΔBMSCs. The cells were cultured in osteogenic media with DMSO, or 100nM, or 300nM IKKiVII for 7 days before harvesting for RNA isolation (n=4). (C) ELISA analysis showing IL-6 secretion from BMSCs isolated from 8-week-old WT (+/+) and Ercc1^−/Δmice. The cells were treated with DMSO or 100nM or 300nM IKKiVII for 3 days. Conditioned media was then harvested for ELISA analysis (n=4). (D) TRAP staining of WT (+/+) and Ercc1^−/ΔBMMs. The cells were cultured in osteoclastogenic media with DMSO or 100nM or 300nM IKKiVII treatment for 6 days prior to TRAP staining (n=5). Scale bar, 50μm. (E) pBMMs-BMSCs co-culture assays. Primary BMSCs from 4-week-old WT (+/+) and Ercc1^−/Δmice were co-cultured with WT (+/+) BMMs for 7–8 days with either DMSO or 100nM or 300nM IKKiVII. The number of TRAP+ MNCs per well was counted (n=5). Scale bar, 100μm. All experiments were performed three times independently, and representative data are shown. All values are shown as mean ± SEM. * p < 0.05 compared to +/+ with DMSO, # p < 0.05 and ## p < 0.01 compared to Ercc1^−/Δwith DMSO.

Fig. 8 — (A) Senescence-associated β-galactosidase staining of WT (+/+) and Ercc1^−/ΔBMSCs that were treated with DMSO (vehicle), or the inhibitor of NF-κB activation IKKiVII (100nM, or 300nM) for 3 days. The percent of positive cells was counted (Right panel, n=4). Scale bar, 100μm. (B) qRT-PCR analysis of expression of osteoblastic markers in WT (+/+) and Ercc1^−/ΔBMSCs. The cells were cultured in osteogenic media with DMSO, or 100nM, or 300nM IKKiVII for 7 days before harvesting for RNA isolation (n=4). (C) ELISA analysis showing IL-6 secretion from BMSCs isolated from 8-week-old WT (+/+) and Ercc1^−/Δmice. The cells were treated with DMSO or 100nM or 300nM IKKiVII for 3 days. Conditioned media was then harvested for ELISA analysis (n=4). (D) TRAP staining of WT (+/+) and Ercc1^−/ΔBMMs. The cells were cultured in osteoclastogenic media with DMSO or 100nM or 300nM IKKiVII treatment for 6 days prior to TRAP staining (n=5). Scale bar, 50μm. (E) pBMMs-BMSCs co-culture assays. Primary BMSCs from 4-week-old WT (+/+) and Ercc1^−/Δmice were co-cultured with WT (+/+) BMMs for 7–8 days with either DMSO or 100nM or 300nM IKKiVII. The number of TRAP+ MNCs per well was counted (n=5). Scale bar, 100μm. All experiments were performed three times independently, and representative data are shown. All values are shown as mean ± SEM. * p < 0.05 compared to +/+ with DMSO, # p < 0.05 and ## p < 0.01 compared to Ercc1^−/Δwith DMSO.