Effect of Insulin receptor deletion in hepatocytes on liver gene expression at different time point in mice either fed ad libitum or upon day-restricted feeding

To assess the consequences of IR deletion on gene expression using microarray and its contribution of IR to the control of rythmic liver gene expression, wild-type (IRhepWT) and KO (IRhepKO) mice were either fed ad libitum or submitted to day-restricted feeding (as described by Damiola et al., 2000). Liver samples were collected around the clock (ZT0, ZT04, ZT08, ZT12, ZT16, ZT20). To generate tamoxifen-inducible hepatocyte-specific IR knockout mouse line (IRhep-/-) and their control (IRhep+/+), animals carrying LoxP sites flanking the fourth exon of the IR gene (IRlox/lox stock number: 006955; Jackson Laboratory, Bar Harbor, ME, USA) were intercrossed with C57BL/6J mice, which specifically express Cre recombinase in the liver under the transthyretin promoter (TTR-CreTam mice), as previously described (Nemazanyy et al., 2015; Smati et al., 2020). Mice were either fed ad libitum (AdLib) or fed only during the day (RF= restricted feeding) for two weeks as previsouly described (Damiola et al., 2000). Total liver RNA was extracted using TRI Reagent (Molecular Research Center, Inc., Cincinnati, OH, USA). Gene expression profiles were obtained at the GeT‐TRiX facility (GenoToul, Genopole Toulouse Midi-Pyrénées) using Agilent Sureprint G3 Mouse GE v2 microarrays (8x60K, design 074809) following the manufacturer's instructions.