Figure 5.

Overexpression of GPX4 decreases PMN transmigration relative to control cells.

A. HCT8 cells were stably transfected with either empty vector pCMV6 or pCMV6-GPX4 and Western blotted for the expression of GPX4. Overexpression of GPX4 resulted in ∼ 30% increase in GPX4 protein expression, and pCMV6 vector also increased GPX4 expression relative to untransfected cells but less so than pCMV6-GPX4 transfected cells (densitometry on right panel).

B. The cells were infected with S. Typhimurium SL1344, followed by PMN transmigration. Cells overexpressing GPX4 show a significant decrease in PMN transmigration compared with untransfected and pCMV6 control vector transfected cells. pCMV6 vector expressing cells which also express relatively higher levels of GPX4 compared with control cells show a significant decrease in PMN transmigration. fMLP was used as a positive control and GPX4 overexpression had no effects on fMLP-induced PMN transmigration (The symbols ‘*’ and ‘**’ indicate statistical significance, using the Student's t-test; P < 0.05, n = 3). Key: white bars = untransfected cells, black bars = pCMV6 vector, grey bars = pCMV6-GPX4