Figure 6.

Isolated cell populations of interest remain viable for cell culture and Next Gen RNA‐sequencing. (A) Fluorescent images of isolated cells at day 12 of cell culture. Cells are stained with fluorescent‐tagged antibodies for EpCAM‐PE (epithelial cell marker), CD49a‐Alexa‐647 (fibroblast marker), and nuclei labeled with hoechst 33342‐blue. (B) dose–response plot shows cell viability, measured by RealTime Glo luminescence, of primary epithelial cells cultured in monoculture and in coculture with patient‐matched primary fibroblasts after 72‐h exposure to docetaxel chemotherapeutic. (C) Volcano plots from cell isolation experiments show average RNA‐seq transcript fold changes across four patients. “EpCAM‐Capture vs. Raw” volcano plot shows transcripts up‐/downregulated in isolated epithelial cells compared to the raw digestate (i), while “CD49a‐Capture vs Raw” shows transcripts up‐/downregulated in isolated stromal cells (ii) with known epithelial and stromal markers labeled with blue dots (orange dots represent transcripts up‐ or downregulated >2‐fold with p‐value > 0.005, green dots represent transcripts up‐ or downregulated > 2‐fold with p‐value < 0.005, red dots represent transcripts up‐ or downregulated with p‐value < 0.005 but by fold change <2, and black dots represent transcripts up‐ or downregulated by fold change <2 and p‐value > 0.005) [Color figure can be viewed at wileyonlinelibrary.com]