Fig. 7: The SLC7A14-induced intestinal sympathetic afferent nerves change is regulated by POMC mTORC1 signaling.
From: Hypothalamic SLC7A14 accounts for aging-reduced lipolysis in white adipose tissue of male mice
a, b P-S6K, t-S6K, p-S6, t-S6, SLC7A14 and TUBULIN proteins in primary hypothalamic cells. c, d mTORC1 kinase assays in primary hypothalamic cells. e IF staining for tdTomato, p-S6 and merge in ARC sections. f IF staining for tdTomato, RAPTOR and merge in ARC sections. g Fat mass by NMR. h Lean mass by NMR. i IF staining for TH. j The NE levels of ileum. k The mRNA levels of Asbt in ileum by RT-PCR. l The TCDCA levels of serum. Studies for (a–d) were conducted using primary hypothalamic cells, infected with adenovirus expressing green fluorescent protein (- Ad-SLC7A14) or Ad-SLC7A14 (+ Ad-SLC7A14) for 48 h in (a) and (c), or transfected with double-stranded siRNA targeting mouse SLC7A14 for 72 h in b and d; e-l were conducted using 5 month-old control mice (- SPKO) or mice with SLC7A14 deletion in POMC neurons (+ SPKO), and injected with AAVs expressing mCherry (- AAV-RAPTOR) or shRAPTOR (+ AAV- shRAPTOR) in (f–l). For a-b, n = 6. For c-d and i, n = 3. For e, n = 6 (- SPKO) and 5 (+SPKO). For (f) n = 5. For (g, h), n = 5 (- SPKO - shRAPTOR), 5 (+SPKO - shRAPTOR), 5 (- SPKO + shRAPTOR) and 6 (+SPKO + shRAPTOR). For (j) and (l) n = 5 (- SPKO - shRAPTOR), 6 (+SPKO - shRAPTOR), 5 (- SPKO + shRAPTOR) and 6 (+SPKO + shRAPTOR). For (k), n = 6 (- SPKO - shRAPTOR), 6 (+SPKO - shRAPTOR), 5 (- SPKO + shRAPTOR) and 5 (+SPKO + shRAPTOR). Data are expressed as the mean ± SEM, with individual data points. Data were analyzed by two-tailed unpaired Student’s t-test (a, b, and e) or ordinary two-way ANOVA with Tukey’s multiple comparisons test (f–h, j–l). Source data are provided as a Source Data file.
