Journal: Physiological Reports

Article Title: The effects of the “living high–training low” model on key proteins involved in cellular aerobic and anaerobic metabolism in male C 57 BL /6 J mice

doi: 10.14814/phy2.70812

Figure Lengend Snippet: Protein content of HIF‐1α in the hypothalamus (panel a), soleus muscle (panel b), white gastrocnemius muscle (panel c) and for nontrained (N) and trained (T) groups kept in normoxia (NOR) and hypoxia (HYP) at the end of study. Panel d shows a representative image confirming the detection of the target protein bands scanned at 800 nm using near‐infrared fluorescence detection. The same membranes were stained for total protein and scanned at 700 nm to normalize for interlane differences in protein loading. The dashed lines in the total protein stain indicate the molecular weight range corresponding to the region in which the target protein bands were detected. Bars are exhibited as M and SEM ( n = 7 per group).

Article Snippet: Regarding primary antibodies, the membranes were incubated with 5% milk in PBS‐Tween for 1 h at room temperature (OXR1, cat# 13514‐1‐AP, Proteintech) or overnight at 4°C (HIF‐1α alpha antibody, cat# NB100‐479, Novus Biologicals and PGC‐1α alpha antibody, cat# NBP1‐04676, Novus Biologicals).

Techniques: Fluorescence, Staining, Molecular Weight

Journal: Physiological Reports

Article Title: The effects of the “living high–training low” model on key proteins involved in cellular aerobic and anaerobic metabolism in male C 57 BL /6 J mice

doi: 10.14814/phy2.70812

Figure Lengend Snippet: Schematic representation of potential physiological interactions between aerobic training and hypoxia discussed in the present study. Although not observed at the time of analysis, it is likely that stabilization of HIF‐1α occurred during the period of hypoxic exposure. Inhibition of prolyl hydroxylase (PHD) activity can enhance anaerobic glycolytic flux and downregulate oxidative metabolism. This metabolic shift may serve as a strategy to reduce reactive oxygen species generation. The LHTL model may have subjected skeletal muscle to heightened stress, as mice exposed to this condition—unlike those trained while living in normoxia—exhibited a pronounced reduction in spontaneous physical activity (SPA) and a diminished ability to complete the prescribed training sessions. While this may appear causal from a statistical viewpoint, it is important not to disregard the possibility of a regulatory central mechanism whereby a signal is sent for the muscle to reduce its activity in order to prevent damage, even in the absence of observable changes in molecular markers typically associated with oxidative stress mitigation, such as PGC‐1α and OXR1.

Article Snippet: Regarding primary antibodies, the membranes were incubated with 5% milk in PBS‐Tween for 1 h at room temperature (OXR1, cat# 13514‐1‐AP, Proteintech) or overnight at 4°C (HIF‐1α alpha antibody, cat# NB100‐479, Novus Biologicals and PGC‐1α alpha antibody, cat# NBP1‐04676, Novus Biologicals).

Techniques: Inhibition, Activity Assay