Anti mir 372

A 10 μl pipette tip was used to generate an artificial wound onto confluent cell monolayers of control or Adeno-SM transduced MDA-MB-231 cells. The cell migration was then observed in serum containing medium upto 24 hours of the above mentioned treatments(s). Images were taken at 37 °C using Motorized IX-81 inverted microscope attached with DP70 CCD camera (Nikon). The distance migrated towards the wound was calculated after 90 minutes. MDA-MB-231 cells, alone or overexpressed with different pCMV-miR constructs or knocked down using antagomiRs in combination with SMAR1 over expression (Adeno-SM) or SMAR1 knockdown (sh1077 construct) were added to the upper chamber of the Boyden chamber (Corning). AntagomiRs used in the assay were anti-miR-371-3p, anti-miR-371-5p, anti-miR-372, anti-miR-373 and anti-miR-negative control (Ambion). The cells migrated to the reverse side of the upper chamber were fixed and stained with Crystal violet and counted under an inverted microscope (Nikon). Data are represented as the average of five fields per treatment well.