Figure 3

Effects of AGR2 ectopic expression on Nthy-ori 3–1 cell growth, migration and invasion. A) Expression levels of AGR2, AGR2 (C → S) and AGR2 (E → A) in transfected Nthy-ori 3–1 cells: after G418 selection, cells were lysed and analyzed by Western blotting with the indicated antibodies. B) Nthy-ori 3–1 cells transfected with AGR2, AGR2 (C → S), AGR2 (E → A) or empty vector (pcDNA) were plated and counted at different time points. Values represent the average of triplicate experiments ± standard deviations. C) A wound was introduced on confluent monolayer of Nthy-ori 3–1 cells transfected with AGR2, AGR2 (C → S), AGR2 (E → A) and vector control (pcDNA) and wound closure was monitored at 12 hours time points. D) Wound closure was measured by calculating pixel densities in the wound area and expressed as percentage of wound closure of triplicate areas ± standard deviations. E) Nthy-ori 3–1 cells transfected with AGR2, AGR2 (C → S) or the empty vector (pcDNA) were seeded in the upper chamber of transwells and incubated for 24 hours; the upper surface of the filter was wiped clean and cells on the lower surface were stained and counted. F) Invasive ability was expressed as number of invaded cells. Values represent the average of triplicate experiments ± standard deviations. Asterisks indicate P < 0.001 (***).