Figure 3.

Intracellular Ca²⁺ chelation prevents shear-induced signaling pathways. Cultured erythroblasts were preincubated for 10 min under static conditions, and subsequently orbitally shaken at 300 rpm for 20 or 60 min (first lane = Time 0). These cultures were left untreated (CTRL, -), treated with solvent DMSO (CTRL, D), or pre-incubated with 1, 3, and 10 μM of the Ca²⁺ chelator BAPTA-AM (BAPTA(μM), 1, 3, 10). Total protein lysates were subjected to SDS-polyacrylamide gel electrophoresis and western blotting. (A) Blots were stained with anti-NFATc2, anti-p-STAT5 and total-STAT5, anti-p-ERK1/2 (Thr202/Tyr204), anti-Total-ERK1/2, anti-p-P38 (Thr180/Tyr182), anti-total p38. (B) Blots were stained with anti-pJNK (Thr183/Tyr185) and anti-Total-JNK.