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. Author manuscript; available in PMC: 2019 Feb 8.
Published in final edited form as: Cell. 2018 Jan 25;172(4):811–824.e14. doi: 10.1016/j.cell.2017.12.038

Figure 7. Differentiation of ADAR1 KO hESCs to NPCs leads to spontaneous IFNβ production, PKR activation, and apoptotic cell death.

Figure 7

(A–D) WT and two independent ADAR1 KO hESC clones were differentiated into monolayer NPCs (Figure S7A). qRT-PCR of IFNβ and two ISGs (STAT1, ISG15) (n = 3 experimental replicates) (A). Western blot of p-PKR in NPCs (B). Bright field images of NPC monolayer at 20 days post differentiation (C). Western blot of full length and cleaved forms of PARP1 and Caspase-3, markers of apoptosis (D). As a positive control for apoptosis, WT NPCs were treated with 1uM of Staurosporine (STA).

(E-F) RIG-I, MDA5, or PKR were knocked down in ADAR1 KO NPCs with lentiviral shRNAs (Figure S7F). Representative western blot (uppler panel) and band intensities normalized to β- actin (lower panels, n = 2~5 experimental replicates) (E). qRT-PCR of IFNβ mRNA (n = 4~5 experimental replicates) (F).

(A, E, F) Data shown as mean ± SEM. Student’s t-test, *P < 0.05, **P < 0.01, ***P < 0.001, ns = not significant.

See also Figure S7.