Figure 4. Correlative studies on pre- and post-biopsy tissues.

A) Example image of standard immunohistochemistry (IHC) analysis for phosphorylated FAK1 (tyrosine 395) from paired patient samples taken pre- and post-treatment. B) Example image of multiplex IHC (left) and quantitation of pFAK high CK19+ cells in partied biopsies C) Patient biopsies were assessed by using flow cytometry (left) or IHC analysis (right) for the presence of different leukocyte subsets. These leukocytes for flow cytometry included the following T cell subsets: total CD8+ cytotoxic lymphocytes (CTLs), Ki67⁺CD8⁺ CTLs, CD4⁺ T effector cells, FOXP3⁺ regulatory T cells, and CD3/CD19⁻CD11b⁺CD14⁺CD68⁺ tumor-associated macrophages. For IHC analysis, total CD8⁺ T cells in the biopsy and CD8⁺ T cells present within 100 μm of CK19+ cells were analyzed by HALO software. All data are displayed as log₂fold changes between pre- and post-biopsies. D) Example IHC analyses for CD8 (brown) and CK19 (red) images for a pair of pre- and post-biopsies from a single patient. *denotes p<0.05 by paired parametric test (B) or Wilcoxon signed-rank test or one-sample test (C) as appropriate.