Supplementary Material from CRISPR/Cas9-Mediated Point Mutation in <i>Nkx3.1</i> Prolongs Protein Half-Life and Reverses Effects <i>Nkx3.1</i> Allelic Loss

Supplementary Tables 1-4: Table 1. Primers for PCR amplification for CRISPR/Cas9 mouse models. Table 2. ssODN oligos. Table 3. Primers for genotyping of mouse mutation. Table 4. Antibodies used for immunofluorescence staining. Supplemental Figures 1-8: Supplementary Figure 1 A. Amino acid sequences of human (top) and murine (bottom) NKX3.1 showing consensus (central) sequence and location of serine 185 and 186 indicated by the box in the figure. B. Effect of S186A mutation on half-life of murine Nkx3.1. Supplementary Figure 2 A. Schematic of targeted sites for mutation at locus. Supplementary Fig 3 A. H&E staining of dorsolateral prostatic lobes at different time points. Results similar to those seen with anterior prostates are shown in the figure. Supplementary Fig 4 A. Immunohistochemistry for cytokeratins 5 and 8 in 2-month old anterior prostate lobes. The arrow in the section from Nkx3.1 show an area of early hyperplasia. The arrows in the section from Nkx3.1 show areas of epithelial detachment. Supplementary Figure 5 Nkx3.1 expression. Supplementary Fig 6 Cell proliferation. Supplementary Figure 7 TUNEL staining was done on paraffin sections from dorsolateral prostate lobes of Nkx3.1 , Nkx3.1 , and Nkx3.1 mice at 2, 6, and 10 months of age. Supplementary Figure 8 A. Cleaved caspase 3 staining was done on paraffin sections from both anterior and dorsolateral prostate lobes of Nkx3.1 , Nkx3.1 , and Nkx3.1 mice at 2, 6, and 10 months of age.