Figure 1.

CAFs stimulate CSC phenotype and CBX4 expression. (A) HSC-5 monolayer cultures maintained in a growth medium were harvested and plated at 4 × 10⁴ cells per well in spheroid growth conditions with normal or CAF-CM and spheroid number monitored for 9 days (left). Representative images on day 9 of growth are shown (right). Scale bars, 200 µm. (B) Spheroid cells were trypsinized and single-cell suspensions were seeded onto Matrigel-coated membranes in Millicell chambers for invasion assays with normal or CAF-CM in the bottom chamber (C) or replated as monolayer cultures and allowed to reach confluence, at which time they were scratched with a 10 ml pipette tip to create a wound; wound closure was monitored over time. (D) After 10 days of spheroid growth in normal or CAF-CM, lysates were electrophoresed for detection of the indicated epitopes.