Fig 2: Immunohistochemical staining for PTK7 in prostate cancer and benign prostate tissue (original magnification ×200). (A) High PTK7 protein expression was found in cytoplasm of prostate cancer tissues; (B) Low PTK7 protein expression was found in cytoplasm of prostate cancer tissues; (C) PTK7 weakly positive staining was found in cytoplasm of benign prostate tissue; and (D) Negative immunostaining of negative controls with the primary antibody omitted in prostate cancer tissues.

Fig 3: Representative western blot analysis of protein tyrosine kinase 7 (PTK7) expression in prostate tissues. The expression of PTK7 protein was significantly increased in prostate cancer tissues compared with benign prostatic hyperplasia tissues. Protein samples obtained from benign prostatic hyperplasia tissues and prostate cancer tissues were analysed using SDS-PAGE followed by immunoblotting with antibody against PTK7. The levels of ß-actin were used as an internal control.

Fig 4: miR-205-5p regulates the gene transcription of PTK7 and suppresses the expression of PTK7 in human colorectal cancer. (A) Binding sites of miR-205-5p (586–593) targeting PTK7-3'UTR-WT and its mutant sequences (PTK7-3'UTR-Mut) are shown. The sites without connections indicate the mutation position. (B) PTK7-3'UTR-WT or PTK7-3'UTR-Mut luciferase reporter was co-transfected into HT29 cells with miR-control, miR-205-5p, inhibitor NC, and miR-205-5p inhibitor, respectively, for 48 h. Luciferase activity of the PTK7 3'UTR was detected using a luciferase reporter gene assay. (C) PTK7-3'UTR-WT or PTK7-3'UTR-Mut luciferase reporters was co-transfected into SW480 cells with miR-control, miR-205-5p, inhibitor NC, and miR-205-5p inhibitor, respectively, for 48 h. Luciferase activity of the PTK7 3'UTR was detected using the luciferase reporter gene assay. (D) Reverse transcription-quantitative polymerase chain reaction analysis of the mRNA expression levels of PTK1 in HT29 and SW480 cells transfected with miR-control, miR-205-5p, inhibitor NC, and miR-205-5p inhibitor, respectively. (E) Protein expression levels of PTK7, measured using western blot analysis, in HT29 and SW480 cells treated as in D. *P<0.05 vs. miR-control. miR, microRNA; PTK7, protein-tyrosine kinase 7; WT, wild-type; Mut, mutant; UTR, untranslated region; NC, negative control.

Fig 5: miR-205-5p accelerates cell proliferation, and inhibits apoptosis through PTK7 in CRC cells. (A) miR-205-5p inhibited the mRNA expression levels of PTK7 in HT29 and SW480 cells. The mRNA expression levels of PTK7 were detected using reverse transcription-quantitative polymerase chain reaction analysis in HT29 and SW480 cells transfected with miR-control, miR-205-5p, miR-205-5p and vector, miR-205-5p and PTK7, respectively (*P<0.05 miR-205-5p, vs. NC; #P<0.05 miR-205-2p+PTK7 vs. miR-205-5p+vector). (B) Western blot analysis was used to measure the protein expression levels of PTK7 in HT29 and SW480 cells treated as in A. (C) An MTT assay was performed to determine the proliferation ability of HT29 cells transfected with miR-control, miR-205-5p, miR-205-5p and vector, and miR-205-5p and PTK7, respectively, at 0, 12, 24 and 48 h *P<0.05 miR-205-5p, vs. NC; #P<0.05 miR-205-2p+PTK7 vs. miR-205-5p+vector. (D) MTT assay was performed to detect proliferation ability of SW480 cells treated as in C. *P<0.05 miR-205-5p, vs. NC; #P<0.05 miR-205-2p+PTK7 vs. miR-205-5p+vector). (E) HT29 and SW480 cells were transfected with miR-control, miR-205-5p, miR-205-5p and vector, and miR-205-5p and PTK7, respectively, and cell apoptosis was detected using Annexin V-FITC/PI staining. *P<0.05 miR-205-5p, vs. NC; #P<0.05 miR-205-2p+PTK7 vs. miR-205-5p+vector. (F) Cell proliferation was detected using colony formation assays in HT29 and SW480 cells transfected with miR-control, miR-205-5p, miR-205-5p and vector, and miR-205-5p and PTK7, respectively. miR, microRNA; PTK7, protein-tyrosine kinase 7; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PI, propidium iodide.