Fig 1. HMB enhances C₂C₁₂ myoblasts cell viability and proliferation.

C₂C₁₂ myoblasts were treated with either 25 μM or 125 μM free acid HMB or vehicle control. Cell viability was assessed at 48 and 72 hours proliferation using the Trypan Blue exclusion assay. Experiments were performed under conditions of both low serum (A) and normal serum (B), (Control, n = 6; 25 μM HMB, n = 3; 125 μM HMB, n = 3, * denotes significant difference compared to control: 3% FBS, P = 0.044; 10% FBS, P = 0.031, One-way ANOVA). C) C₂C₁₂ myoblasts were treated with 25 μM or125 μM HMB or vehicle control for 48 hours in proliferation media (10% FBS) at which point total viable cells were counted while gating out cellular debris with the Scepter™ Automated Cell Counter (Control, n = 6; 25 μM HMB, n = 3; 125μM HMB, n = 3,* denotes significance compared to control: 25 μM HMB, P<0.042; 125 μM HMB, P<0.01, One-way ANOVA).