Fig. 6
From: Glucose promotes cell growth by suppressing branched-chain amino acid degradation
BCAA accumulation is required for mTOR activation during cardiomyocyte growth. a Cellular BCAA level was measured in NRCMs treated with phenylephrine (PE, 100 μM) or vehicle at indicated time points (*p < 0.05 vs. Vehicle, n = 5). b Cellular BCAA level was examined in NRCMs transduced with indicated adenovirus and treated with phenylephrine (PE, 100 μM) or vehicle for 3 h (*p < 0.05 vs. LacZ/Vehicle, #p < 0.05 vs. LacZ/PE, n = 5). c NRCMs transduced with indicated adenovirus were treated with phenylephrine (PE, 100 μM) or vehicle for 6 h. Immunoblots of cell lysates (left) and statistical analyses of densitomeric measurements of p-p70 S6K (Thr 389) and p-mTOR (Ser 2448) (right) are shown (*p < 0.05 vs. sh-con/LacZ/Vehicle, #p < 0.05 vs. sh-con/LacZ/PE, &p < 0.05 vs. sh-con/KLF15/PE, n = 4). SE for short exposure. d, e NRCMs transduced with indicated adenovirus were incubated with DMEM containing either glucose or non-glucose substrates for one hour and then treated with phenylephrine (PE, 100 μM) or vehicle. d Immunoblots of cell lysates (left) and statistical analyses of densitomeric measurements of p-p70 S6K (Thr 389) and p-mTOR (Ser 2448) (right) are shown (*p < 0.05 vs. sh-con/Glucose/Vehicle, #p < 0.05 vs. sh-con/Glucose/PE, &p < 0.05 vs. sh-con/Lactate/PE, n = 4). e 48 h after PE treatment, myocytes were fixed and stained with anti-Troponin T. Cell surface area in each group was quantified and expressed relative to the control (*p < 0.05 vs. sh-con/Glucose/Vehicle, #p < 0.05 vs. sh-con/Glucose/PE, &p < 0.05 vs. sh-con/Lactate/PE, n = 4). Scale bar, 25 μm. Data shown as mean ± s.e.m. P values were determined using one-way ANOVA followed by Newman-Keuls comparison test (a, b, c, d) or Kruskal–Wallis test followed by Dunn’s comparison test (e)
