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-Thr^37/46-4E-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.