Figure 2.

Effect of heterozygous Spink1 deletion (Spink1-KO^het) on mRNA and protein expression in the mouse pancreas. (A) CRISPR/Cas9-mediated genome engineering was used to delete the entire Spink1 gene. E, Exons; F and R, forward and reverse genotyping primers; gRNA, guide RNAs. (B) Expression of Spink1 mRNA in the pancreas of C57BL/6N and Spink1-KO^het mice was quantified by reverse-transcription quantitative PCR and expressed relative to the average value of the C57BL/6N strain. (C) Protein levels of SPINK1 in pancreas homogenates were determined by Western blotting and densitometry. (D) Protein levels of mouse cationic trypsinogen (T7) were assessed by Western blotting and densitometry. On Western blots, ERK1/2 was measured as loading control. Representative blots from 2 experiments are shown. Densitometry results are expressed as percent of the average band intensity of the C57BL/6N strain. Individual data points with mean and standard deviation are shown. The difference of means between the groups was analyzed by 2-tailed unpaired t-test.