Generation and Characterization of Breast Cancer PDXs

Breast cancer PDXs were generated from one luminal (HCI-011) and four TN (HCI-001, HCI-002, HCI-004, HCI- 010) breast cancers serially passaged in NOD/SCID mice as described previously50 (link)51 (link). Tumour fragments were placed into cleared inguinal fat pads of pre-pubescent NOD/SCID mice, grown to 20–25 mm and subsequently dissected and stored by freezing in 90% FBS and 10% dimethylsulfoxide until used or fixed in paraffin for IHC. Total RNA was extracted according to the manufacturers' instructions using RNeasy Plus Mini kit (Qiagen, Hilden, MD, USA) and iScript Reverse Transcription Supermix for RT-QPCR (BIO-RAD) was used for cDNA synthesis. RT-QPCR was performed using iTAG Universal SYBR Green Supermix (BIO-RAD).