*4.12. Proliferation Assay*

The proliferation rate of 12Z cells 72 h after LINC01133 knockdown was analyzed using the CyQuant direct cell proliferation Assay (Invitrogen, Waltham, MA, USA) according to the manufacture's protocol. Prior to the assay, transfected cells were trypsinized 48 h after transfection with targeting and control siRNA oligos, seeded on 96 flat-bottom cell culture plates (Thermo Fisher Scientific, Roskilde, Denmark) at a concentration of 15,000 cells/well and allowed to grow for an additional 12 h. The level of fluorescence was accessed with the Clariostarplus microplate reader (BMG Labtech, Ortenberg, Germany) with filters appropriate for 480 nm excitation and 520 nm emission maxima. The observed fluorescence values were first corrected for the background fluorescence determined with a cell- free sample, and a standard curve was used to estimate cell number. All measurements were performed in technical triplicates, and the average number of proliferating cells relative to control siRNA-treated cells was set to 1.
