Figure 13.

RNA sequencing and representative images of IHC conducted on lung sections harvested from the orthotopic injection mice model. (A) A bubble plot showing the top 10 enriched Hallmark pathways under PTK7-enriched conditions as determined by GSEA analysis. (B) Representative images of immunocytochemistry (ICC) targeting SLUG and ZEB1 in PLC/PRF/5 cells sorted by their PTK7 expression. 4′,6-Diamidino-2-phenylindole (DAPI) staining was used to locate the nucleus of stained cells. The average nuclear localization of the SLUG and ZEB1 was quantified by ImageJ and compared between PTK7^-ve and PTK7^+ve cells by the Student t test. (C) Representative images of ICC targeting SLUG in MHCC97L, of which PTK7 was overexpressed and subsequently suppressed by lentiviral-based knockdown using 2 independent short hairpin RNA structures (sh845 and sh923). DAPI staining was used to locate the nucleus of stained cells. The average nuclear localization of SLUG was quantified by ImageJ and compared between the 4 transfected lines by 1-way analysis of variance with multiple t tests. (D) Images showing IHC staining of PTK7 and SLUG performed on lung metastases resulted from orthotopic liver injection of MHCC97L cells with or without PTK7 overexpression. DN, down-regulated; E2F, E2F transcription factor; FDR, false discovery rate; G2M, G2 to M phase cell cycle transition; IF, immunofluorecence; IL, interleukin; NTC, nontarget control; OE, overexpression; STAT5, signal transducer and activator of transcription 5. P values (∗P < .05, ∗∗P < .01, ∗∗∗P < .001).