Fig. 8: POMC SLC7A14 regulates mTORC1 signaling via TSC1.
From: Hypothalamic SLC7A14 accounts for aging-reduced lipolysis in white adipose tissue of male mice
a, b P-TSC1, t-TSC1, SLC7A14 and TUBULIN proteins in primary hypothalamic cells. c P-S6K, t-S6K, p-S6, t-S6, t-TSC1, SLC7A14 and TUBULIN proteins in primary hypothalamic cells. d P-TSC1, t-TSC1, IKKβ, SLC7A14 and TUBULIN proteins in primary hypothalamic cells. e, g-h Immunoblotting and co-immunoprecipitation in primary hypothalamic cells. f Venn chart of interactions for SLC7A14, IKKβ, and TSC1 in HitPredict. i Working model. Studies for (a–e, g) and h 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, or transfected with double-stranded siRNA targeting mouse SLC7A14 for 72 h in b, or transfected with double-stranded shRNA targeting mouse TSC1 (+ Sh-TSC1) or without TSC1 shRNA (- Sh-TSC1) for 72 h and infected with adenovirus expressing green fluorescent protein (- Ad-SLC7A14) or Ad-SLC7A14 (+ Ad-SLC7A14) for 48 h in (c), or infected with adenovirus expressing green fluorescent protein (- Ad-SLC7A14) or Ad-SLC7A14 (+ Ad-SLC7A14) and transfected with (+ IKKβ−3xflag) or without (- IKKβ−3xflag) FLAG-tagged IKKβ plasmids for 48 h in d; or infected with adenovirus expressing green fluorescent protein (- Ad-SLC7A14) or Ad-SLC7A14 (+ Ad-SLC7A14) for 48 h. Immunoprecipitation (IP) and immunoblotting (IB) were performed using the antibodies indicated in (e–h). For (a-d) and (g), n = 6. For e and h, n = 3. Data are expressed as the mean ± SEM, with individual data points. Data were analyzed by two-tailed unpaired Student’s t-test (a and b), or ordinary two-way ANOVA with Tukey’s multiple comparisons test (c, d). Source data are provided as a Source Data file.
