IHC Analysis of Subcutaneous Tumor Samples

The collected subcutaneous tumors were fixed with 4% formalin, embedded in paraffin and then sectioned into 4-μm-slices. The standard protocol for IHC of the TMAs and subcutaneous tumors using a streptavidin-peroxidase (SP) Kit (Zhongshan biotech, Beijing, China), as described previously [25 (link)]. The slides were incubated with antibodies specific for TFAP2C (Abcam, cambridge, MA, USA), TRIM29 (Proteintech, Chicago, IL, USA), PHLPP1 (Proteintech, Chicago, IL, USA), p-AKT (Cell Signaling Technology, Danvers, MA, USA) and Ki-67 (Cell Signaling Technology, Danvers, MA, USA). The images of IHC staining were obtained by an Olympus microscope (Tokyo, Japan).