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They are smaller, too, which may indicate some metabolic downmodulation, although mitochondrial mass and transmembrane potential are not notably altered in them, suggesting that mitochondrially mediated cell size determining factors may not be causal [ 46, 47].
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We found that TSC2's potential to counteract mTORC2-mediated cell size control indeed depends on functional Rheb.
Two-dimensional contour blot analyses of FSC versus DNA content demonstrated that mTORC2-mediated cell size regulation is not a consequence of its effects on cell cycle progression described above.
To clarify the role of this pathway for mTORC2-mediated cell size and cell cycle regulation, we first wanted to investigate the effects of rictor-specific siRNA treatment in cells with knockdown of TSC2.
Last but not least, to test whether Akt regulation indeed represents the initial relevant step for the mTORC2-mediated cell size control, we tested the effects of rictor-specific siRNA treatment in cells expressing myristoylated Akt.
Genetic epistasis results fit well into a model where dSesn-mediated cell size suppression occurs between AKT and p70S6K, where the AMPK-TSC2 axis acts; as reducing expression of TSC1, TSC2 or AMPK hampered dSesn-dependent cell growth inhibition (Lee et al, 2010).
However, this is unlikely to be the whole story because in cells where Cdc2 Tyr15 phosphorylation is prevented from occurring (thus bypassing Pom1-mediated regulation) cell size homeostasis is maintained, albeit with a broader size distribution [ 50].
Thus, cell-cycle arrest and mitochondria-mediated increases in cell size are distinct, and LipidMix supplementation does not inhibit cell-size increase but acts by stimulating cell proliferation.
Further assessment of cellular hypertrophy indicated that DON inhibition of HBP flux also abolished the Ad-LXRα-mediated reductions in cell size that was increased upon PE stimulation (Fig 6G and H).
To investigate whether TSC2 is essentially involved in the mTORC2-mediated cell cycle and/or cell size regulation, we analysed the effects of TSC2-specific siRNA treatment on the rictor-siRNA-induced deregulations described above.
Our results suggest a novel role for CYP2S1-mediated metabolism in modulating BEAS-2B cell size.
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