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. Author manuscript; available in PMC: 2023 Nov 1.
Published in final edited form as: Nat Cell Biol. 2023 Apr 27;25(5):643–657. doi: 10.1038/s41556-023-01131-x

Extended Data Fig. 5. Esrrb KO clones characterisation.

Extended Data Fig. 5

a: Left: Bar plot showing the number of AP positive colonies after clonal assay of cells with loxP sites flanking the second exon of both alleles of Esrrb (Esrrb fl/fl, dark blue) and Esrrb KO cells generated by Cre-mediated recombination (light blue), cultured in 2iL and differentiated for 24h, 48h and 72h in N2B27. Mean +/- SD of n=3 independent experiments, shown as dots. Right: Barplots showing expression measured by qPCR of Esrrb in Esrrb fl/fl (dark blue) and Esrrb KO (light blue) cells cultured in 2iL and differentiated for 24h, 48h and 72h. Mean of n=2 independent experiments.

b: Schematic representation of edited alleles of 3 CRISPR-generated Esrrb KO clones. The edited genome is indicated in red. The blue sequence is an insertion. Black bars indicate deletions.

c: Bright field images of 3 CRISPR-generated Esrrb KO clones, cultured in 2iL and after 48h of 2iL withdrawal. Scale bar: 300μm.

d: Barplot showing expression by RNAseq of naive markers in WT E14 cells and in 3 CRISPR-generated Esrrb KO clones. WT values were set as 1. Mean of n=2 biological replicates.