Journal: EBioMedicine

Article Title: Clock Gene Dysregulation Induced by Chronic ER Stress Disrupts β-cell Function

doi: 10.1016/j.ebiom.2017.03.040

Figure Lengend Snippet: Attenuation of DBP transcriptional activity induced by chronic ER stress. (a) The Dbp and Nfil3/E4bp4 mRNA expression levels in Wfs1 +/+ a/a, Wfs1 +/+ A y /a , Wfs1 −/− a/a and Wfs1 −/− A y /a islets at ZT12. Data are presented as relative expressions, and each point represents the mean + SEM (N = 3/group; ★★ p < 0.01). (b–c) MIN6 cells (b) and mouse islets (c) were incubated with thapsigargin at the indicated concentrations for 24 h and then subjected to real-time RT-PCR. Data are expressed relative to the values for cells incubated without thapsigargin and are the means + SEM of six experiments. ★ p < 0.05, ★★ p < 0.01. (d) Mouse islets were incubated with tunicamycin at the indicated concentrations for 24 h and then subjected to real-time RT-PCR. Data are expressed relative to the values for cells incubated without tunicamycin and are the means + SEM of six experiments. ★★ p < 0.01.

Article Snippet: Normal rabbit IgG (sc-2027; Santa Cruz), rabbit anti-RNA polymerase II (Millipore), rabbit anti-DBP antibody (AVIVA SYSTEM BIOLOGY), and anti-E4BP4 (sc-28,203; Santa Cruz) were used for immunoprecipitation.

Techniques: Activity Assay, Expressing, Incubation, Quantitative RT-PCR

Journal: EBioMedicine

Article Title: Clock Gene Dysregulation Induced by Chronic ER Stress Disrupts β-cell Function

doi: 10.1016/j.ebiom.2017.03.040

Figure Lengend Snippet: DBP and E4BP4 directly regulate the transcription of genes linked to insulin secretion. (a) Real-time PCR analysis of clock-controlled genes, Slc2a2 , Ins1 , Ins2 and Rab37, in isolated islets during a 12 h light-dark cycle (N = 6 per time point). Data are means ± SEM. ★ p < 0.05, ★★ p < 0.01 (controls versus TG-C). (b) E4BP4 at the selected gene promoters in pancreatic islets isolated from TG-C mice. UCSC genome browser images of E4BP4 profiles at Ins1 , Ins2 , Slc2a2 and Rab37 in TG-C mice. Chromosome coordinates are indicated at the top. The plot in the middle shows the density of ChIP-seq reads with the peak score indicated on the Y-axis. The Ins1 , Ins2 , Slc2a2 and Rab37 genes are shown at the bottom. Red scale bars indicate 1 kb of genome. (c) Schematic models depicting the circadian control of insulin release via regulation of the basal ATP/ADP ratio and the gene expressions of Slc2a2 , Ins1 , Ins2 and Rab37 in islets.

Article Snippet: Normal rabbit IgG (sc-2027; Santa Cruz), rabbit anti-RNA polymerase II (Millipore), rabbit anti-DBP antibody (AVIVA SYSTEM BIOLOGY), and anti-E4BP4 (sc-28,203; Santa Cruz) were used for immunoprecipitation.

Techniques: Real-time Polymerase Chain Reaction, Isolation, ChIP-sequencing