Fig. 13

Validation of relative gene expression levels in vitro for genes included in the APPCAFRS. The relative expression levels of THBS2 (A), COL5A1 (B), MARCKS (C), and DPT (D) in WPMY-1 cells and CAFs. The relative expression levels of THBS2, COL5A1, MARCKS, and DPT in pre-ADT and post-ADT CAFs (E). An example of a T-cell early activation assay. CD4 + T cells that were cocultured with different CAF subtypes. After exclusion by forward- and side-scatter, residual viable cells were stained for CD4, CD25 and CD69. A representative example of CD4 gating is shown in the left Panel (F). Each of the other panels shows the CD69 + population upon coculture of T cells with pre-ADT CAFs or post-ADT CAFs following incubation with OVA (G, H)