Figure 1. Repeated transfection of hESCs with p53-CRISPR/Cas9 shows efficient downregulation of p53 protein while maintaining pluripotency.

A) Scheme of repeated transfection using Neon Transfection System. B) Analyses of p53 protein expression by Western blot after 2-5 consecutive rounds of transfection. Alpha-tubulin was used as loading control. C) Relative expression of miR-34a, or miR-302a (D), as determined by qPCR. Protein levels of pluripotency markers (Oct4, Nanog; E) and differentiation-associated markers (GATA6, Brachyury; F) were analyzed in MOCK and p53 Low hESCs by Western blotting. Differentiating hESCs were used as positive control. Alpha-tubulin was used as loading control. Mean data ± SEM from at least three independent experiments are shown. (*) denotes p ≤ 0.05.