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. 2017 May 9;8:15142. doi: 10.1038/ncomms15142

Figure 1. Generation and in vitro characterization of a Phf8 null allele.

Figure 1

(a) Diagram of the Phf8locus around exons 7 and 8 for wild type (WT) (top), targeted (middle) and knockout (KO) (bottom) alleles. (b) Southern blot analysis confirms deletion of exons 7 and 8 of Phf8 in embryonic stem cells (ESCs) using StuI digest and probe A (see panel A). (c) Western blot analysis confirms loss of Phf8 protein in KO ESCs. (d) RNA-seq tracks from Phf8 KO and WT ESCs show absence of reads for deleted exons 7 and 8 in KO sample. (e) Western blot analysis detecting indicated histone modifications in Phf8 KO ESCs. (f) Growth curve for Phf8 WT and KO ESCs transduced with either empty vector control (EV) or full length Phf8 cDNA (Phf8) lentiviral vectors. Error bars=s.e.m. *=p<0.05 for WT EV and KO EV. ŧ=p<0.05 for KO Phf8 and KO EV (Student's t-test, two-tailed). (g) Representative images of Phf8 WT and KO neurosphere cultures grown for 1 week from single cell suspensions. Scale bar: 750 μm (h) Quantification of neurosphere diameter 1 week after plating of single cell suspension. *=p<0.05, ***=p<0.001 (Student's t-test, two-tailed).