Figure 10.

PTK7 in vitro functional assays performed on various HCC cell lines. (A) PTK7 was overexpressed in MHCC97L cells and subsequently suppressed by lentiviral-based knockdown using 2 independent short hairpin RNA (shRNA) structures (sh845 and sh923). The number of cells migrating or invading in Transwell assays were compared with EV + nontarget control (NTC) by 1-way analysis of variance with multiple t tests. The successful knockdown of PTK7 was validated by Western blot. (B) Representative images and the comparison of the number of foci formed by sorted PLC cells and MHCC97L cells overexpressing PTK7. Statistical comparisons were performed by the Student t test. (C) The predicted tumor-initiating cell frequency of sorted PLC cells and MHCC97L overexpression cells was determined by extreme limiting dilution assay and analyzed with a publicly available algorithm. The frequency was represented by the estimated population of cells required for the detection of 1 tumor-initiating cell. (D) Representative H&E staining of a lung tissue section harvested from mice injected with RIL175 through the tail vein. Metastases are indicated with black arrows. Ctrl, control; OE, overexpression. P values (∗P < .05, ∗∗P < .01, ∗∗∗P < .001).