Figure 5. In LMX1A-knockout AGS cells altering KCNQ1OT1 expression fails to change cell viability and proliferation. AGS cells were transfected with the lentiCRISPR/Cas9 LMX1A knockout constructs containing non-overlapping sgRNA sequences (“S1/S2”), following FACS sorting and puromycin selection two stable lines were obtained (“Cas9-LMX1A-ko”). LMX1A and LMX1B protein expression was tested (A, LMX1A protein results were quantified). LV-KCNQ1OT1 or KCNQ1OT1 siRNA (500 nM) were transduced to the Cas9-LMX1A-ko AGS cells (“S1/S2”) for 72h, KCNQ1OT1 expression (B), cell viability (C) and proliferation (D) were shown. (E) The proposed signaling cascade of this study. For each assay, n=5. *P <0.05 vs. “Ctrl” cells. Experiments in this figure were repeated three times, and similar results were obtained.