Extended Data Fig. 7: Tsc2S1365A/S1365A knock-in mice display significantly increased mTORC1 but no change in mTORC2 activation; depressed autophagy in Tsc2S1365A/S1365A mice subjected to pressure overload is reversed by mTOR inhibition with everolimus.
From: PKG1-modified TSC2 regulates mTORC1 activity to counter adverse cardiac stress
a, Immunoblots and summary quantification for mTORC2 targets from Tsc2WT/WT and Tsc2S1365A/S1365A mice subjected to sham or pressure-overload surgeries and treated with sildenafil or vehicle. n = 4 biologically independent experiments, box and whisker plots (median) with individual data are shown; data normalized to median of Tsc2WT/WT sham. One-way ANOVA with Tukey multiple comparisons test. P70S6K is shown at the top as an mTORC1 control, showing increased phosphorylation with pressure overload that is greater and unresponsive to sildenafil in Tsc2S1365A/S1365A mice. *P = 0.002 versus sham, †P = 0.04 versus pressure overload, ‡P = 0.03 versus sham. However, there were no significant changes (P ≥ 0.62 between conditions within genotype) in the expression of mTORC2 substrates: phosphorylated (S473)/total AKT, phosphorylated (T24/T32)/total FOXO1/3 and phosphorylated(T346)/total NRDG1. b, LC3-II expression is unaltered while p62 expression increases from pressure overload in Tsc2S1365A/S1365A myocardium, indicating suppression of autophagy. Both are reversed by treatment with the mTORC1 inhibitor everolimus. n = 6 biologically independent animal experiments, data are mean ± s.d., one-way ANOVA with Tukey multiple comparisons test, *P ≤ 3 × 10−5 versus other two groups. Data are normalized to the mean of sham control.
