TCHQ-induced, ROS-mediated, prolonged p-ERK expression in splenocytes.

<p>(A) Freshly isolated splenocytes were treated with various concentrations of PCP or TCHQ for 2 hr. The total protein lysates were extracted to analyze the expression of p-ERK and ERK by western blot. β-actin was served as a loading control. N: normal group; D: DMSO control group. The protein expression for the indicated periods of time is presented as (B). (C) The splenocytes were pretreated with or without 5 mM NAC for 1 hr. After washing with ice-cold PBS, the cells were treated with 100 µM PCP or 25 µM TCHQ for 15 min or 2 hr. the Total protein lysates were extracted to analyze the expression of p-ERK and ERK by western blot. ERK served as a loading control. (D) The cells were pretreated with U0126 (10 µM) for 30 min followed by treatment of 25 µM TCHQ for 2 hr. Total cell lysates were prepared and immunoblotted using antibodies to cleaved caspase-3, PARP, p-ERK, ERK and β-actin. β-actin was served as a loading control. Statistical analysis was performed using a two-tailed Student’s <i>t</i>-test. *, p<0.05.