Transient ER-stress increases PLD3 expression during myogenesis.

<p>(<b>A</b>) Western blot analysis of PLD3 in differentiating C2C12 myoblasts pre-treated for 30 min with thapsigargin (TG, 1 µM), tunicamycin (TN, 2 µg/ml), Brefeldin-A (BFA, 2 µM) or DMSO before differentiation for the indicated periods of time. (<b>B</b>) Proliferating C2C12 myoblasts were treated with TG, TN, BFA, or DMSO for the indicated time followed by western blot analysis for PLD3 expression. (<b>C</b>) Western blot analysis of HeLa cells pre-treated with TN without (D0) and with (subsequent days) serum reduction for the indicated times. (<b>D</b>) Western blot analysis of C2C12 myoblasts pre-treated as shown with DMSO, TN, 1 mM DTT, or DTT and 10 µM calcium ionophore A23187 before differentiation for 3 days. The PLD3 and GAPDH levels of expression were determined using Licor odyssey software. The PLD3 level of expression was normalized to the level of GDPDH expression and is presented as fold-change with respect to the DMSO control. Representatives of three independent experiments are shown.