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. 2020 Apr 6;16(11):1861–1875. doi: 10.7150/ijbs.45112

Figure 3.

Figure 3

Osblr8 is required for the maintenance of pluripotency. A) Osblr8 knockdown in iPSCs. After lentiviral shRNA transfection and puromycin selection, iPSCs colonies were selected and expanded for Q-PCR. shCT: random shRNA control; shOsblr8-1, -2: Lenti Osblr8 shRNA-1 and Osblr8 shRNA-2 vectors. For comparison, the abundance of Osblr8 in iPSCs was set as 1. ** p<0.001 as compared with iPSCs and random shCT controls. B) Osblr8 knockdown downregulates stem cell core factors in iPSCs. * p<0.05 as compared with iPSC and random shCT controls. C) Effects of Osblr8 on iPSCs proliferation. MTT assay was performed to determine the viability of iPSCs transfected with shOsblr8, shCT and untreated iPSCs, respectively. D) Osblr8 knockdown induces iPSC differentiation. The Osblr8 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 OCT4 (red). Compared to the control group, shOsblr8 transfected iPSCs were negative for OCT4 immunostaining and were differentiated morphologically (yellow dotted line).