Electrogenetic cellular insulin release for real-time glycemic control in type 1 diabetic mice

Data underlying the figures in the publication “Electrogenetic cellular insulin release for real-time glycemic control in type 1 diabetic mice”, published in <em>Science</em>, <strong>2020</strong>, 368, 993-1001. https://science.sciencemag.org/content/368/6494/993 Table of contents: <strong>1. Electrogenetics source data file</strong>; File openable with Graphpad Prism containing the source data for the main <em>Figures: 1-4 </em>(Transgene expression by SEAP measurement), <em>5b, 6a, 6f</em> (Insulin by ELISA), <em>5c, 6b, 6e </em>(NanoLuc luciferase), 7c (NanoLuc luciferase in vivo) and <em>7a, 7d</em> (Glycemia). <strong>Transgene expression by SEAP measurement</strong> Data for main <em>Figures 1, 2, 3, 4</em>. SEAP (human placental secreted alkaline phosphatase) levels were profiled in cell culture supernatants using a colorimetric assay. 100 µL 2x SEAP assay buffer (20 mM homoarginine, 1 mM MgCl2, 21% diethanolamine, pH 9.8) was mixed with 80 µL heat-inactivated (30 min at 65°C) cell culture supernatant. After the addition of 20 µL substrate solution (120 mM p-nitrophenyl phosphate; cat. no. AC128860100, Thermo Fisher Scientific), the absorbance time course was recorded for 45 min at 405 nm and 37°C using a Tecan Genios PRO plate reader (cat. no. P97084; Tecan Group AG, Maennedorf, Switzerland) and the SEAP levels were determined as follows: first, absorbance change over time (slope) was calculated. According to the Beer–Lambert’s law, absorbance is proportional to the concentration of a colored compound and depends on the light path length (d) and extinction coefficient (ε) (ε for p-nitrophenyl (εpNP) = 18.600 M−1 cm−1). Enzymatic activity EA [U/L] was calculated from the equation: EA = slope × dilution factor × εpNP−1 × d−1 Values in the file present determined SEAP levels. <strong>Insulin by ELISA</strong> Data for <em>Figures 5b, 6a, 6f</em>. Values in the file present Insulin level as determined by ELISA kit. The assay was performed according to manufacturer’s instructions. <strong>NanoLuc luciferase </strong> Data for <em>Figures 5c, 6b, 6e</em>. NanoLuc® luciferase was quantified in cell culture supernatants using the Nano-Glo® Luciferase Assay System (cat. no. N1110; Promega, Duebendorf, Switzerland). In brief, 7.5 µL of cell culture supernatant was added per well of a black 384-well plate and mixed with 7.5 µL substrate-containing assay buffer. Total luminescence was quantified using a Tecan Genios PRO plate reader (Tecan Group AG). Values in the file present measured luminescence levels. <strong>NanoLuc luciferase in vivo </strong> Data for <em>Figure 7c.</em> Aliquots of 15 µL of whole-blood samples were diluted in 5 μL of 50 mM EDTA and frozen at -20 °C until NanoLuc® quantification as described above. Values in the file present measured luminescence levels normalized to time point 0 (Normalization individually for each mouse). <strong>Glycemia </strong> Data for <em>Figures 7a, 7d</em>. Blood glucose level was determined using a glucometer (Contour®Next, Bayer Healthcare, Leverkusen, Germany). Values in the file present measured glucose levels. <strong>2. Figure 7b</strong>; Data for <em>Figure 7b</em>. File openable with Graphpad Prism. Blood glucose level was determined using a glucometer (Contour®Next, Bayer Healthcare, Leverkusen, Germany). Values in the file present measured glucose levels. <strong>3. Figure 6c and 6d</strong>; Excel file with the data for <em>Figures 6c, 6d</em>. NanoLuc® luciferase was quantified in cell culture supernatants using the Nano-Glo® Luciferase Assay System (cat. no. N1110; Promega, Duebendorf, Switzerland). In brief, 7.5 µL of cell culture supernatant was added per well of a black 384-well plate and mixed with 7.5 µL substrate-containing assay buffer. Total luminescence was quantified using a Tecan Genios PRO plate reader (Tecan Group AG). Values in the file present measured luminescence levels.