Supplementary Figure 5: Construction of PAX5 knockout hESC line with CRISPR.
From: A PAX5–OCT4–PRDM1 developmental switch specifies human primordial germ cells
(a) Targeting strategy of PAX5 knockout in hESC with the designated guide RNA (gRNA) and the resulting deleted sequences. (b) Sequences of wide type PAX5 and PAX5 KO line that show homologous recombination and deletions are shown. Grey box indicates CRISPR recognition site and black bars indicate deleted sequences. (c) Immunofluorescence of PAX5 on wild-type (WT) and PAX5 KO after BMPs-induced differentiation. Scale bars represent 50 µm. Immunostaining experiments were independently repeated a minimum of three times with similar results. (d) Western blot of PAX5 and ACTIN on wild type (WT) and PAX5 knockout (KO) cells. Western blot images are representative of three independent experiments. Unprocessed scans of Western blot analysis are available in Supplementary Fig. 8.
