Fig. 1: Generation of Haspin-KO cell lines through CRISPR/Cas9. | Cell Death Discovery

Fig. 1: Generation of Haspin-KO cell lines through CRISPR/Cas9.

From: Haspin balances the ratio of asymmetric cell division through Wnt5a and regulates cell fate decisions in mouse embryonic stem cells

Fig. 1: Generation of Haspin-KO cell lines through CRISPR/Cas9.

A Schematic illustration of the creation of Haspin-KO mESCs through CRISPR/Cas9 technology. The blue sequences are gRNA sequences, and the red sequences are PAM sequences. F and R primers were used to perform genomic PCR. B Genomic PCR for identifying Haspin knockout. C qRT‒PCR analysis of Haspin expression levels in WT, Haspin-KO, and rescued cells. D Western blotting analysis of H3T3ph and H3S10ph in WT, Haspin-KO, and rescued cells. E The statistical results of protein expression in (D) (n = 3). F Immunofluorescence of H3T3ph and H3S10ph in WT, Haspin-KO, and Haspin-eGFP overexpressing (rescue) cells. Scale bar: 20 μm. G The statistical results of the fluorescence intensity of (F). n = 30 from three independent experiments. The data in (C), (E), and (G) are represented as the mean ± SEM. *p < 0.05, **p < 0.01 and ***p < 0.001.

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