Figure 6.

TNFR1 knockdown inhibits BV6-induced cell elongation, migration and invasion. (a) T98G cells were lentivirally transduced with shRNA against TNFR1 or vector control and knockdown of TNFR1 was assessed by western blotting. GAPDH expression served as loading control. (b–d) T98G cells transduced with shRNA against TNFR1 or control vector were treated for 24 h with 2.5 μM BV6. Cell morphology was analyzed by phase-contrast microscope. Cell elongation was quantified by measuring cell length and width and by calculating cell elongation index (length/width); fold increase in elongation index is shown (b). Migrated cells were fixed and stained with crystal violet. Migration was quantified by dissolution of crystal violet staining and determination of optical density (c). Cells were allowed to invade in a collagen-coated transwell migration chamber; invasive cells were fixed, stained with crystal violet and optical density was measured (d). In (a), a representative experiment of two independent experiments is shown, in (b–d) mean+S.D. values of three independent experiments performed in triplicate (b) or duplicate (c and d) are shown; *P<0.05; n.s. not significant