Crossbreeding Protocol for Mouse Models

The SHP2-floxed mice (25 (link)) were obtained from Dr. Benjamin Neel, the HER2/Neu transgenic mice (11 (link)) were purchased from the Jackson Laboratory, and the MMTV-Cre (26 (link)) were obtained from Dr. Timothy Lane. A standard crossbreeding protocol as outlined in Figure 1a was used in the study. Genomic DNA isolated from tail snips was used for PCR genotyping of mice. The forward and the reverse primers in a respective order include 5’-TAGCTGCTTTAACCCTCTGTGT-3’ and 5’-CATCAGADCAGGCCATATTCC-3’ for SHP2, 5’-TTTCCTGCAGCAGCCTACGC-3’ and 5’CGGAACCCACATCAGGCC-3’ for HER2, and 5’-ATCCGAAAAGAAAACGTTGA-3’ and 5’-ATCCAGCTTACGGATATAGT-3’ for Cre. The internal control primers for the HER2 transgene were the forward 5’-CAAATGTTGCTTGTCTGGTG-3’ and the reverse 5’-GTCAGTCGAGTGCACAGTTT-3’. All genetic studies were conducted by strictly following the approved protocol by the West Virginia University Institutional Committee for animal Use and Care (WVU ICUC).