Phospho-antibody microarray analyses for islets of control and IFT88 knockout mice [PEX100]

β-cell specific IFT88 knock-out mice recapitulate human diabetes with impaired insulin secretion and altered islet hormone paracrine regulation. To examine the signaling pathways regulating islet cell function, we subjected protein lysates of whole islets from control and IFT88 knockout mice to a commercial phospho-antibody array analysis (Full Moon Bio, Inc). Samples were probed against 1318 site-specific and phospho-specific antibodies with 2 replicates per antibody on 76 x 25 x 1mm glass slides. IFT88 fl/fl mice and INS1-Cre mice on a C57BL/6 background were crossed to generate β-cell specific IFT88 knockout mice. Animals were weaned at age 3 weeks and maintained on a standard chow diet. Islets were isolated using modified Lacy protocol and allowed to recover in culture overnight before collection and lysis. Total protein lysates were prepared from 240 size-matched islets per sample (2 controls and 2 knockouts, each pooled from 2 mice) and used for FullMoonBio Phospho Explorer Array (PEX100) analysis.