Research Paper Volume 11, Issue 11 pp 3731—3749

Nectandrin B-mediated activation of the AMPK pathway prevents cellular senescence in human diploid fibroblasts by reducing intracellular ROS levels

Figure 5. Effect of NecB on the expression and/or activation of sirtuins, protein synthesis regulators, and cell survival/death-related proteins. Young and old HDFs (Y and O) were treated with vehicle or 10 or 20 μg/mL NecB (N10 or N20 in old HDFs) for 2 days, The cell lysates were analyzed by western blot with antibodies for sirtuins (A), such as SIRT1−7, protein synthesis regulators (B), including P-Thr2446-mTOR, mTOR, raptor, P-Thr389-p70S6K, p70S6K, P-Thr37/46-4E-BP1, and 4E-BP1, and cell survival/death-related proteins (C), including P-Tyr458-p85, p85 (PI3K), P-Ser473-Akt, Akt, P-Tyr204-ERK1/2, ERK1/2, P-Thr85-p38, p38, P-Thr183Tyr185-JNK, JNK1/2, P-Ser83-ASK1 (inactive form), P-Thr845-ASK1 (active form), and total ASK1. β-actin was used as an internal control.