Fig. 2.

CD167a expression enhances cell migration in vitro. a Hierarchical clustering of COL1A1, COL1A2 (collagen genes), and CD167a (collagen receptor) genes expression in a human bladder cancer patient cohort (cohort III: TCGA); red and green colors indicate high and low expression, respectively. Grey box indicates patients with co-expression of COL1A1/COL1A2 and CD167a genes. b Immunohistochemical analyses of collagen I and CD167a in representative human MIBC tissues verified the localization of CD167a positive cancer cells in adjacent to stromal collagen I expression. Scale bar:100 μm. c Left panel: Western blot analyzing CD167a protein expression in mCherry-CBLuc-Control and mCherry-CBLuc-CD167a-T24 cancer cells. Middle panel: Representative images of mCherry-CBLuc-Control and mCherry-CBLuc-CD167a-T24 cancer cell migration capacity in vitro. Right panel: Quantification of percent migration at 10 h post-wound induction relative to 0 h. d, e Combinatorial effects of exogenous collagen I and CD167a overexpression in cancer cell migration in vitro. Doxycycline-inducible CD167a-expressing T24 cancer cells were subjected to the wound-healing assay with or without collagen I treatment. Cell lysates were harvested after collagen I and doxycycline (15 ng/ml) stimulation for subsequent western blot evaluation at the indicated time points (0, 6, and 18 h). Left panel: Representative images of wound closure at 0 and 10 h upon treatment with 25 μg ml⁻¹ collagen I. Right panel: Quantification of percent migration at 10 h post-wound induction relative to 0 h. Statistical analysis: a two-tailed, unpaired student’s t-test. Error bar: mean ± SEM, n = 3 independent experiments. ^*p < 0.05, ^**p < 0.01