Investigating miR-584-5p Effects on Clonogenic and Migration Abilities

MB cells were transfected with miR-584-5p, miR-NC, scramble-siRNA, or target-siRNA (Sigma) for 24 h, harvested, and subjected to long-term clonogenic and migration assays^{62 (link)}. For the long-term clonogenic assays, 1000 cells/well were reseeded in six-well plates for an additional 7–10 days until colonies were visible. Colonies were fixed with 4% paraformaldehyde and stained with 1% crystal violet. For the transwell migration assay, 100,000 cells were reseeded in Corning BioCoat Control Cell Culture Inserts with 8.0-µm PET Membrane in serum-free medium. Cells were allowed to migrate toward complete media for 24 h before fixation with 4% paraformaldehyde and staining with 1% crystal violet.