Fig. 5. Specially inhibition of ATF4 in amygdalar PKC-δ neurons blocks leucine deprivation-induced WAT browning.

a Gene expression of Atf4 and Trb3 in amygdala by RT-PCR. b Daily food intake. c Fat mass by NMR. d Subcutaneous WAT (sWAT) weight. e Representative images of hematoxylin and eosin (H&E) staining of sWAT. f sWAT cell size quantified by Image J analysis of H&E images. g Gene expression of Ucp1, Pgc1a, Cidea, Dio2, and Prdm16 in sWAT by RT-PCR. h Representative images of immunohistochemistry (IHC) of UCP1 in sWAT. i UCP1 protein in sWAT by western blotting (left) and quantified by densitometric analysis (right); A.U.: arbitrary units. Studies for a were conducted using 14- to 15-week-old male wild-type mice fed a control (Control) or leucine-deficient [(-) L] diet for 3 days; studies for (b–i) were conducted using 13- to 16-week-old male PKC-δ-Cre mice receiving AAVs expressing mCherry (PKC δ − DN ATF4) or DN ATF4 (PKC δ + DN ATF4) fed a Control or (-) L diet for 3 days. Data are expressed as the mean ± SEM (n represents number of samples and are indicated above the bar graph), with individual data points. Data were analyzed by two-tailed unpaired Student’s t test for (a), or one-way ANOVA followed by the SNK (Student–Newman–Keuls) test for (b–i). Source data are provided as a Source data file.