HCC cells (5 × 104) were added in 24-well plates followed by transfection of indicated RNA constructs or controls. After 1, 2, and 3 days absorbance was measured at 450 nm via Multiskan GO (Thermo Fisher Scientific, USA).
When EdU uptake was being assessed 24 hours after transfection with appropriate mimics, inhibitors, or control constructs, HCC cells were labeled with EdU and imaged using the BeyoClick™ EdU-555 Imaging Kit (Beyotime, Beijing, China). DAPI was used to stain nuclei in this experiment, and the number of EdU+ cells was determined as a readout for proliferation.
When EdU uptake was being assessed 24 hours after transfection with appropriate mimics, inhibitors, or control constructs, HCC cells were labeled with EdU and imaged using the BeyoClick™ EdU-555 Imaging Kit (Beyotime, Beijing, China). DAPI was used to stain nuclei in this experiment, and the number of EdU+ cells was determined as a readout for proliferation.