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. 2020 May 1;318(6):H1487–H1508. doi: 10.1152/ajpheart.00709.2019

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Fig. 7. — Effects of BMAL1 deletion and/or SR-9009 administration on parameters involved in cardiac glycogen metabolism. A: cardiomyocyte-specific BMAL1-knockout (CBK) and littermate control (CON) mice challenged with either SR-9009 (SR; 100 mg·kg⁻¹·day⁻¹ ip) or vehicle (Veh) at zeitgeber time (ZT)9 for 8 consecutive days, followed by heart isolation 6 h after the last administration (i.e., ZT15) and subsequent assessment of glycogen content. B: hearts were isolated from CBK and littermate control (CON) mice at ZT0, ZT3, ZT6, ZT12, ZT15, ZT18, and ZT21 (ZT0 and ZT24 are identical time points and data; data are duplicated only for illustration purposes, but not for statistical analysis), followed by assessment of glycogen content. C: CBK and littermate control (CON) mice challenged with either SR (100 mg·kg⁻¹·day⁻¹ ip) or vehicle (Veh) at ZT9 for 8 consecutive days, followed by heart isolation 6 h after the last administration (i.e., ZT15) and subsequent assessment of protein expression by Western blotting (normalized to amido black staining): phosphorylated glycogen synthase (pGS; i and ii), total glycogen synthase (GS; i and iii), and the phospho-to-total ratio (pGS/GS; iv). D: samples from C used for assessment of glycogen phosphorylase (i and ii). E: samples from C used for assessment of phosphorylated Akt (p-Akt; i and ii), total Akt (Akt; i and iii), and the phospho-to-total ratio (p-Akt/Akt; iv). F: samples from C used for assessment of phosphorylated mTOR (p-mTOR; i and ii), total mTOR (mTOR; i and iii), and the phospho-to-total ratio (p-mTOR/mTOR. G: samples from C used for assessment of phosphorylated GSK3β (p-GSK3β; i and ii), total GSK3β (GSK3β; i and iii), and the phospho-to-total ratio (p-GSK3β/GSK3β; iv). H: samples from C used for assessment of LC3II (i and ii). I: samples from C used for assessment of p62 (i and ii). J: samples from C used for assessment of STBD1 (i and ii). Data are reported as means ± SE for 11–13 mice/experimental group. Main effects for SR-9009 and/or genotype are reported at the top of each graph.

Fig. 7. — Effects of BMAL1 deletion and/or SR-9009 administration on parameters involved in cardiac glycogen metabolism. A: cardiomyocyte-specific BMAL1-knockout (CBK) and littermate control (CON) mice challenged with either SR-9009 (SR; 100 mg·kg⁻¹·day⁻¹ ip) or vehicle (Veh) at zeitgeber time (ZT)9 for 8 consecutive days, followed by heart isolation 6 h after the last administration (i.e., ZT15) and subsequent assessment of glycogen content. B: hearts were isolated from CBK and littermate control (CON) mice at ZT0, ZT3, ZT6, ZT12, ZT15, ZT18, and ZT21 (ZT0 and ZT24 are identical time points and data; data are duplicated only for illustration purposes, but not for statistical analysis), followed by assessment of glycogen content. C: CBK and littermate control (CON) mice challenged with either SR (100 mg·kg⁻¹·day⁻¹ ip) or vehicle (Veh) at ZT9 for 8 consecutive days, followed by heart isolation 6 h after the last administration (i.e., ZT15) and subsequent assessment of protein expression by Western blotting (normalized to amido black staining): phosphorylated glycogen synthase (pGS; i and ii), total glycogen synthase (GS; i and iii), and the phospho-to-total ratio (pGS/GS; iv). D: samples from C used for assessment of glycogen phosphorylase (i and ii). E: samples from C used for assessment of phosphorylated Akt (p-Akt; i and ii), total Akt (Akt; i and iii), and the phospho-to-total ratio (p-Akt/Akt; iv). F: samples from C used for assessment of phosphorylated mTOR (p-mTOR; i and ii), total mTOR (mTOR; i and iii), and the phospho-to-total ratio (p-mTOR/mTOR. G: samples from C used for assessment of phosphorylated GSK3β (p-GSK3β; i and ii), total GSK3β (GSK3β; i and iii), and the phospho-to-total ratio (p-GSK3β/GSK3β; iv). H: samples from C used for assessment of LC3II (i and ii). I: samples from C used for assessment of p62 (i and ii). J: samples from C used for assessment of STBD1 (i and ii). Data are reported as means ± SE for 11–13 mice/experimental group. Main effects for SR-9009 and/or genotype are reported at the top of each graph.