Figure 4.

Capsaicin enhances TRIB3 mRNA expression and protein stability.

Notes: (A) AGS, MKN-45, and SiHa cells were treated with 10 µM MG132 for the indicated time periods, cell lysates were prepared, and the protein levels of TRIB3 were detected by immunoblotting. (B) Cells were treated with 50 µg/mL CHX for the indicated time periods, cell lysates were prepared, and the protein levels of TRIB3 were measured. (C) AGS cells were exposed to capsaicin for 24 hours, total mRNA was isolated, and real-time PCR was used to measure the mRNA expression of TRIB3. n=5. *P<0.05. The results of each experiment were normalized with respect to the expression of the internal control gene, TBP. (D) AGS and (E) MKN-45 cells were pretreated with or without capsaicin for 22 hours and then co-treated with 50 µg/mL CHX for 0–2 hours. The protein levels of TRIB3 were determined. All data were obtained from at least three to six independent experiments. n=3 for AGS, n=6 for MKN-45.

Abbreviations: CHX, cycloheximide; PCR, polymerase chain reaction; TBP, TATA- box binding protein.