Peblr20 lncRNA is required for pluripotent state maintenance. (A)
Peblr20 knockdown in iPSCs. After lentiviral shRNA transfection and puromycin selection, iPSCs colonies were selected, expanded and collected for Q-PCR. shCT: random shRNA control; shPeblr20: Peblr20 shRNA. For comparison, the abundance of Peblr20 in iPSCs was set as 1. ** p<0.01 as compared with iPSCs and random shCT controls. (B)
Peblr20 knockdown downregulates stem cell core factors in iPSCs. * p<0.05 as compared with iPSC and random shCT controls. NS: not significant. (C)
Peblr20 knockdown induces iPSC differentiation. The Obelr20 shRNA and random shRNA lentiviral vectors carry the copGFP reporter gene (green). After lentiviral transfection, cells were fixed and immunostained by an antibody against the stem cell pluripotent marker NANOG (red). Compared to the control group, shPeblr20 transfected iPSCs were negative for NANOG immunostaining and were differentiated morphologically (blue dotted line).