FIG 4.
Ectopic expression of NleF upregulates IL-8 expression. (A) IL-8 expression levels of HeLa cells 24 h after transfection with pRK5::myc-gfp, pRK5::myc-nleF, or pRK5::myc-nleF1–185. Cells were either left unstimulated or stimulated with 20 ng/ml TNF for 16 h. mRNA levels for IL-8 were analyzed by RT-qPCR. IL-8 expression levels were significantly higher in the TNF-treated control and NleF-transfected cells than in GFP-transfected cells. Results are averages for at least 3 independent biological repeats. Asterisks indicate significant differences (P < 0.05) from results for GFP-transfected cells. (B) Levels of Myc-NleF or Myc-NleF1–185 were analyzed in whole-cell extracts by Western blotting 24 h posttransfection. Similar levels of NleF and NleF1–185 were observed relative to tubulin levels. Results are representative of two independent biological repeats.