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. 2020 Jan 28;20:62. doi: 10.1186/s12885-019-6435-1

Fig. 8.

Fig. 8

Effect of NFкB (RelA) knockdown on migration, invasion and clonogenicity of U87MG cells. Representative images (20x magnification) and quantification of (a) cell migration and (b) cell invasion in U87MG after NFкB (RelA) knockdown are shown. Transwell cell migration and invasion (matrigel) assays were performed for 24 and 48 h respectively, using U87MG cells harvested after 48 h of siRelA/siControl transfection. Cells that migrated across transwell membrane or invaded through matrigel were fixed, stained and counted. Mean of five non-overlapping fields was calculated in each of the triplicate wells. Number of cells that migrated/invaded in siRelA cells were normalized against the number of cells that migrated/invaded in siControl cells. Values in graphs are expressed as fold change. Significant decrease in migration and invasion was observed in siRelA-treated U87MG cells as compared to the siControl cells as determined by Student’s t-test. c Anchorage-independent colony formation assay. Representative micrographs (4x magnification; scale bar: 50 μm) and quantification of colonies in soft agar assay of U87MG after NFкB (RelA) knockdown are shown. U87MG cells were harvested after 48 h of siRelA/siControl transfection and 15,000 cells/well were seeded in 0.3% agarose in DMEM supplemented with 20% FBS in triplicate wells in a six-well plate. After 22 days, colonies were stained with crystal violet. Mean number of colonies (≥30 μm size) of ten non-overlapping fields was calculated in each of the triplicate wells in siRelA/siControl cells. Graph values are expressed as mean ± s.d. Significant difference was observed in siRelA-treated U87MG cells as compared to the siControl cells as determined by Student’s t-test