Figure 2.

Generation and characterization of HCC1806-RR in vitro. HCC1806-RR line was generated using the SB transposon system by a co-transfection of transposon plasmids encoding for dual expression of RFP and Rluc or a puromycin resistance gene in combination with SB transposase. (A, B) Morphology of expanded monoclonal cell populations was compared to the parental cell line by bright-field microscopy. (C) Monoclonal isolates with identical morphology were then assessed for homogenous RFP expression using fluorescent microscopy. (D) Average RLU/s per cell was calculated to ensure a linear correlation with luciferase activity measured in triplicate and expressed as the mean RLU/s per cell ± SD. (E) All selected clones were assessed for unaltered growth pattern as compared with the parental cell line. Each time point was performed in triplicate, repeated twice, and presented as the mean cell number ± SD. Student's t test analysis showed no statistical difference between the proliferation rates of the HCC1806-RR clone and the parental line.