Dri chem7000

Serum levels of 1,25 (OH)₂ vitamin D₃ were measured with an EIA kit by immunoextraction followed by quantitation with an enzyme-immunoassay (ImmunoDiagnosticSystems PLC, Tyne and Wear, UK; Cat# AC-62F1). Mouse serum FGF23 was measured using the Quidel ELISA Kit that measures C-terminal FGF-23 according to the manufacturer’s instructions (Quidel, San Diego, USA; Cat# 60–6300). Mouse PTH serum levels were determined with an ELISA Kit that measures intact PTH (PTH 1–84) (Quidel, San Diego, USA; Cat 60–2305). Mouse serum chemistries were determined with a Dri-Chem7000 chemistry analyzer (Heska). Urine chemistries were determined with a Roche ModP analyzer. Ionized serum calcium measurements were performed with a Siemens RAPIDlab 348EX machine.

Blood was collected via cardiac puncture and the serum was isolated and stored at −80°C. Serum aliquots were used to measure liver alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels using a DRI-CHEM 7000 (HESKA, Loveland, CO). Representative data from three independent experiments are shown. n=3-4 per group, per experiment. Data are presented as mean unit/liter (U/L) ± SEM.

The generation of Egf^−/− mice and Tgfa^−/− mice has previously been reported (5 (link)). We utilized Egf^−/WTTgfa^−/WT mice that were on a mixed genetic background to produce the following experimental groups: WT mice, Egf^−/− mice, Tgfa^−/− mice, and Egf^−/−Tgfa^−/WT mice. These mice were obtained from MMRRC and showed a substantial degree of heterozygosity (Mouse Universal Genotyping Array [MUGA] showed that ~30% of the genome was heterozygous; C57bl6 and Sv129 mixed genetic background). Mouse serum BUN levels and chemistries were determined with a Dri-Chem7000 chemistry analyzer (Heska). Urine microalbumin was measured with a Siemens DCA Vantage analyzer. The exact number of mice for which urinary microalbumin, serum BUN levels, and serum chemistries were measured are indicated in Figures 1B, 1C, and S2.