In the present study, we observed increased levels of p-ERK
and p-P38, and a lower level of p-JNK in cells treated with 10 [micro]M MEHP.
In addition, since, according to our current and previous studies, photoreceptor cell apoptosis induced by light injury peaked in 1 day, it is better to set more time points before the conclusion of this 24h period to observe the progressive process of p-ERK
elevation and cell apoptosis.
In our study, we found that LPS single treatment significantly up-regulated the expressions of I[kappa]B[alpha], p65, p-JNK, p-ERK
, and p-p38 in RPTECs, and BAI co-treatment obviously alleviated these increases.
LPS exposure to RAW 264.7 cells resulted in significant activation in MAPKs including p-ERK
, p-P38, and p-JNK, whereas the LPS-induced activation of ERK was prominently blocked by both P.
The protein expression levels of Erk, p-Erk
, JNK, p-JNK, p38 and p-p38 were determined by western blot and quantification analysis.
(a) Western blot and analysis of P38, p-P38, JNK, p- JNK, ERK, and p-ERK
. (b) Western blot and analysis of p-MKK3/6, MKK3 and MKK6.
Levels of MAPK proteins (p38, ERK, and JNK) and their phosphorylated form (p-p38, p-ERK
, and p-JNK, resp.) were measured as mean fluorescence intensity (MFI) by flow cytometry at different time points after AGE stimulation.
PVDF membranes were blocked with 5% skim milk at room temperature for 1 h to avoid nonspecific binding, and immunoblots were incubated for overnight at 4[degrees]C with primary antibodies that specifically detect I[kappa]B[alpha], p-[kappa]B[alpha], JNK, p-JNK, p38, p-p38, ERK, p-ERK
, COX-2, iNOS, and [beta]-tubulin.
(d) The expressions of PTEN, [alpha]-SMA, actin, p-AKT, AKT, p-ERK
, and ERK by Western blot in fibroblasts with or without okadaic acid treatment on FC.
The protein expressions of ZO-1, occludin, TLR4, MyD88,p-JNK, JNK, p-ERK
, and ERK in the gut were detected by Western blotting.
10, UVB dramatically initiated the phosphorylation of ERK (p-ERK
) and p38 (p-p38) and FV quenched UVB-induced p-ERK
Nevertheless, the mRNA and protein expression of p-ERK
and CHOP were all significantly increased by the TM treatment, in a dose independent manner, but this effect was suppressed by HQH (p < 0.01; Figure 4B, 4C, and 4D).