Figure 6. Development of acute iron overload (IO) in vivo model and validation of IO and insulin resistance in skeletal muscle.

• A
  Schematic diagram of iron‐dextran injections’ experimental plan.
• B
  Prussian Perl blue staining in liver and skeletal muscle after iron injections.
• C, D
  Representative Western blot images and quantification of ferritin and Tfr1 (indicated by arrowhead) to tubulin in skeletal muscles 24 h after iron injections. *P < 0.05 (unpaired Student's t‐test versus control).
• E
  Insulin tolerance test 24 h after iron injections.
• F
  Glucose tolerance test (GTT) 24 h after iron injections.
• G
  Quantification of area under curve in ITT (E). *P < 0.05 (unpaired Student's t‐test versus control).
• H
  Quantification of area under curve for panel (F). *P < 0.05 (unpaired Student's t‐test versus control).
• I, J
  Representative Western blot images and quantification of phospho‐IRS1 (Y612) and phospho‐AKT (T308) to GAPDH in skeletal muscles 24 h after iron injection followed by i.p. insulin injection. *P < 0.05 (unpaired Student's t‐test versus control).

Data information: Results are presented as mean ± SEM. n = 6 males for ITT and GTT. n = 3 males for Western blot analysis. Scale bar = 50 μm.