Fig. 1: Chronic rapamycin suppresses mTORC1 and mTORC2 signaling in cultured Tsc1-cKO neurons.
From: Raptor downregulation rescues neuronal phenotypes in mouse models of Tuberous Sclerosis Complex
a Schematic of the experiment, created with BioRender.com. b Representative western blots (WB) from Tsc1fl/fl cultures with (+) or without (−) four-day rapamycin (Rapa) treatment. MW indicates molecular weight. WT=Tsc1fl/fl + AAV-GFP; KO=Tsc1fl/fl + AAV-Cre-GFP. Two independent samples per genotype are shown. This experiment was replicated 5 times. c–h Bar graphs display WB quantification (mean ± SEM) for the indicated proteins, expressed as a percentage of Control (WT) levels. Phospho-proteins were normalized to their respective total proteins. Dots represent data from individual culture wells. WT n = 11, KO n = 12, WT+Rapa n = 13 and KO+Rapa n = 13 culture wells from 5 independent culture preps; 2 mice per culture. ns = non-significant. Dashed lines at 100% indicate Control levels. All statistical tests were two-sided and P values were corrected for multiple comparisons. c Tsc1, Kruskal–Wallis, p < 0.0001; WT vs KO, ****p < 0.0001; WT vs WT+Rapa, p > 0.9999; WT vs KO+Rapa, ****p < 0.0001; KO vs KO+Rapa, p > 0.9999; Dunn’s multiple comparisons tests. d Raptor, One-way ANOVA, p = 0.0001, F(3, 45) = 8.495; WT vs KO, *p = 0.0221; WT vs WT+Rapa, p = 0.2187; WT vs KO+Rapa, p > 0.9999; KO vs KO+Rapa, *p = 0.0161; Sidak’s multiple comparisons tests. e Rictor, One-way ANOVA, p < 0.0001, F(3, 45) = 12.09; WT vs KO p = 0.4323; WT vs WT+Rapa, **p = 0.0010; WT vs KO+Rapa, p = 0.1773; KO vs KO+Rapa, **p = 0.0025, Sidak’s multiple comparisons tests. f p-S6 Ser240/244, One-way ANOVA, p < 0.0001, F(3, 45) = 41.21; WT vs KO, ****p < 0.0001; WT vs WT+Rapa, **p = 0.0014; WT vs KO+Rapa, **p = 0.0015; KO vs KO+Rapa, ****p < 0.0001; Sidak’s multiple comparisons tests. g p-4EBP1 T37, One-way ANOVA, p < 0.0001, F(3, 45) = 9.997; WT vs KO, p = 0.1355; WT vs WT+Rapa, ****p < 0.0001; WT vs KO+Rapa, ***p = 0.0003; KO vs KO+Rapa, p = 0.1218; Sidak’s multiple comparisons tests. h p-Akt Ser473, One-way ANOVA, p < 0.0001, F(3, 45) = 107.6; WT vs KO, ****p < 0.0001; WT vs WT+Rapa, ****p < 0.0001; WT vs KO+Rapa, ****p < 0.0001; KO vs KO+Rapa, ****p < 0.0001; Sidak’s multiple comparisons tests. See also Supplementary Figs. 1, 2. Source data are provided as a Source Data file.
