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Alkaline phosphatase staining

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  • SEM and TEM were used to evaluate the surface morphology and microstructure of the fiber membrane. Ultraviolet spectrophotometry was used to detect drug release. A LIVE/DEAD Viability/Cytotoxicity Kit and fluorescence staining were used to detect cell morphology and activity. Alkaline phosphatase and calcium mineralization deposition were used to evaluate the osteoinductive activity of the scaffold. Dynamic mechanical analysis was used to determine the Young’s modulus, maximum load, and maximum elongation of the prepared scaffold.

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  • In the current study, the effect of USP7 on pluripotency maintenance of mouse ESCs (mESCs) has been investigated with the help of cell viability assay, morphological analysis, alkaline phosphatase (ALP) staining, qPCR analysis, and Western Blotting. 600 nM P5091 application, which showed no significant toxicity in mESCs, increased the total ubiquitinated protein amount as a proof of the accomplishment of proper USP7 inhibition.

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  • Monoclonal antibodies directed against PSA clone 28A4 (Novacastro, UK) in a concentration of 2,5μg/ml were used to detect prostate cells, and identified using a detection system based on alkaline phosphatase-antialkaline phosphatase (LSAB2 DAKO, USA) with new-fuschin as the chromogen. To permit the rapid identification of positive cells there was no counter staining with Mayer´s hematoxilin. Levisamole (DAKO, USA) was used as an inhibitor of endogenous alkaline phosphatase, with positive and negative controls.

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