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Correction: Alpl prevents bone ageing sensitivity by specifically regulating senescence and differentiation in mesenchymal stem cells

机译:校正:ALPL通过特异性调节间充质干细胞中的衰老和分化来防止骨老化敏感性

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摘要

a SA-β-gal staining and Ki67, γH2AX and LAP2β immunostaining of third-passage MSCs from normal controls and HPP patients. Quantification of Ki67+, γH2AX+ and LAP2β+ is indicated in the right panel. Scale bars: 50 μm. b Expression levels of ageing-specific genes in normal and HPP yiMSCs were examined by western blotting. Scale bars, 50 μm. c Expression levels of CD73 and CD39 in normal and HPP MSCs were examined by western blotting. d Extracellular ATP concentrations in normal and HPP MSC medium were examined by a regular ATP concentration assay. e Intracellular radioactivity was examined after a 1-h treatment with ATP-γ-32P in different lentiviral vector transduction groups. f Intracellular ATP concentrations were assayed 48 h after the transduction of different lentiviral vectors. g Western blotting analysis of p-AMPKα expression in normal control and the ALPL shRNA, HPP control and pLenti-ALPL groups. h Expression levels of p16 and p53 were assayed 48 h after the transduction of different lentiviral vectors. i HPP MSCs overexpressing ALPL or treatment with 0.1 mmol·L−1 metformin, Alizarin Red staining and quantification of mineralized nodules were performed on day 28 after osteogenic induction (OS). Expression levels of Runx2 and OCN were examined by western blotting on day 7 after induction. j Oil Red O staining and quantification of fat depots were performed on day 14 after the adipogenic induction (AD). PPAR-γ expression was examined on day 7 after induction by western blotting. Scale bars, 100 μm. (N) Normal control n = 5, HPP (hypophosphatasia patient) n = 2. The data are presented as the means ± s.d. of each independent experiment performed in triplicate. *P < 0.05, **P < 0.01. e–f, i–j One-way analysis of variance (ANOVA). a, d Unpaired two-tailed Student’s t-test.
机译:来自正常对照和HPP患者的第三段MSCs的SA-β-加仑染色和KI67,γH2AX和LAP2β免疫染色。右侧面板表示Ki67 +,γH2AX+和LAP2β+的定量。秤条:50μm。 B Western印迹检查正常和HPP yimscs中老化特异性基因的B表达水平。秤条,50μm。蛋白质印迹检查CD73和CD39中CD73和CD39的C表达水平。通过常规ATP浓度测定检查正常和HPP MSC培养基中的细胞外ATP浓度。在用不同的慢病毒载体转导组中用ATP-γ-32P处理1小时处理后检查细胞内放射性。在转导的不同慢病毒载体的转导之后测定细胞内ATP浓度48小时。 G Western印迹分析正常对照中P-AMPKα表达及ALPL shRNA,HPP控制和最新铝族基团。在转导后48小时的不同慢病毒载体进行测定,P16和P53的H表达水平。 I HPP MSCs过表达ALPL或用0.1mmol·L-1二甲双胍治疗,在成骨诱导后的第28天进行茜素红染色和矿化结节的定量,在第28天进行(OS)。诱导后第7天通过Western印迹检查Runx2和OCN的表达水平。 J油红色O染色和脂肪库的定量在脂肪诱导(AD)后第14天进行。 Western印迹诱导后第7天检查PPAR-γ表达。秤条,100μm。 (n)正常对照n = 5,HPP(次磷酸患者)n = 2。数据被呈现为平均值±S.D.每个独立的实验一式三份。 * P <0.05,** P <0.01。 E-F,I-J单向分析方差(ANOVA)。 A,D未配对的双尾学生的T检验。

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