RD and K562 cells were transfected with either siRNAs or LNA-modified oligonucleotides as previously described. 24 h after transfection, an equal number of transfected live cells were plated into a 12-well plate. At the same time, the indicated concentrations of either Vincristine (Selleckchem), Taxol (Paclitaxel - Selleckchem) or Nocodazole (Sigma) were added to the medium.
SK-N-BE cells were transfected with either si-SCR or si-Circ as previously described.
24 h after transfection cells were plated in 96-well plates (3x104 cells/well), and the following day treated with the indicated concentrations of either Taxol or Vincristine.
24 h after the addition of the drugs, viable RD and SK-N-BE cells were stained with crystal violet solution (0.5%) and then the absorbance was read at 600 nm, using a Glomax-Multi+ Detection System machine (Promega). Instead, dead/live K562 cells were counted with the addition of 0.4% Trypan Blue stain (ThermoFisher Scientific) at a CytoSmart cell counter (Corning). Survival was expressed as percentage of cells alive normalized to DMSO-treated cells (vehicle control), and untreated cells were used to control each condition.
SK-N-BE cells were transfected with either si-SCR or si-Circ as previously described.
24 h after transfection cells were plated in 96-well plates (3x104 cells/well), and the following day treated with the indicated concentrations of either Taxol or Vincristine.
24 h after the addition of the drugs, viable RD and SK-N-BE cells were stained with crystal violet solution (0.5%) and then the absorbance was read at 600 nm, using a Glomax-Multi+ Detection System machine (Promega). Instead, dead/live K562 cells were counted with the addition of 0.4% Trypan Blue stain (ThermoFisher Scientific) at a CytoSmart cell counter (Corning). Survival was expressed as percentage of cells alive normalized to DMSO-treated cells (vehicle control), and untreated cells were used to control each condition.