Fb12 luminometer

The assay was performed as previously described 35. Briefly, pGL3‐ARE 36 or pGL3‐DRE 37, 38 and pRL‐TK (thymidine kinase promoter‐driven renilla luciferase plasmid) were transiently cotransfected into cells. After 6 hrs, medium was replaced with fresh medium and 10% CDFBS. Cells were then cultured for 48 hrs with or without DHT (10 nM). After 24 hrs, cells were washed with 1xPBS and then incubated in the presence of 100 μl CCLR (cell culture lysis reagent) (Promega, WI, USA) at room temperature for 30 min. Cell lysates were then placed in a microtube and centrifuged at 12000 g for 5 min. Supernatant (5 μl) was then mixed with 50 μl luciferase assay reagent. Luciferase activity was measured immediately using a luminescence reader (Berthold Detection System FB12 Luminometer) and presented as relative luminescence units.

GOx, WS₂, HRP, benzoquinone, tert-butyl alcohol, and TMB were acquired from Sigma Co. (St. Louis, MO, USA, www.sigmaaldrich.com). Rhodamine B (RB), NaOH, H₂O₂, glucose, and SDS were obtained from Merck Co. (Darmstadt, Germany, www.merck.com). All chemicals were obtained in analytical grade and used without any treatment. All experimental solutions were prepared by double-deionized (DI) water. The morphological assessments of synthesized nanosheets were carried out with a JEOL HR-TEM (JEM-2200FS, acting at 200 kV, Tokyo, Japan, www.jeol.co.jp). XRD patterns were acquired using a Siemens D5000 X-ray diffractometer (Berlin, Germany, www.siemens.com) with a Cu Kα exciting source (λ = 1.54056 Å). Florescence analyses were performed using an LS-45 PerkinElmer spectrofluorometer (Waltham, MA, USA, www.perkinelmer.com). An S2000 spectrophotometer (WPA Lightwave, Cambridge, England, www.biochrom.co.uk) was used to obtain UV–visible absorption data. An IviumStat potentiostat/galvanostat (Eindhoven, The Netherlands, www.ivium.nl) was utilized for electrochemical investigations. CL experiments were done using a Berthold FB12 luminometer (Bad Wildbad, Germany, www.berthold.com).