Fig. 2

Subcellular localization of S. pyogenes Cas9 tagged with various nuclear localization sequences. a Table of non-native NLS and NES sequences tested; basic residues are underlined for NLSs and hydrophobic residues are underlined for NESs. b Fluorescence microscopy of live yeast cells containing NLS sequences (GFY-3435 to 3437) or NES sequences (GFY-3438, 3439) fused to eGFP-tagged Cas9 (also see Table 1). Yeast were cultured in galactose prior to imaging. An integrated copy of Nup188-mCherry marked the nuclear periphery. Representative images are shown; white dotted lines, outline of selected cells. Scale bar, 3 μm. Triangles indicate the yeast vacuole