Fig 1: Overexpression of GAB1 reverses the inhibitory effect of PTK6 interference on the invasion and migration abilities of cervical cancer cells. (A) Representative images of wound healing assay (magnification, ×100) and quantification of cell migration. (B) Representative images of Transwell assay (magnification, ×100) and quantification of cell invasion. (C) RT-qPCR and (D) western blotting were used to assess MMP12 and MMP9 expression levels. *P<0.05 and ***P<0.001 vs. Control or siRNA-PTK6 + Ov-NC. GAB1, GRB2-associated binding 1; PTK6, protein tyrosine kinase 6; RT-qPCR, reverse transcription-quantitative PCR; MMP, matrix metalloproteinase; siRNA, small interfering RNA; Ov, overexpression; NC, negative control.
Fig 2: Overexpression of GAB1 reverses the inhibitory effect of PTK6 interference on the proliferation of cervical cancer cells. (A) RT-qPCR and (B) western blotting respectively, detected GAB1 expression in cervical cancer C-33A cells. ***P<0.001 vs. Ov-NC. (C) C-33A cell proliferation was detected by CCK-8 assays. ***P<0.001 vs. Control; ##P<0.01 and ###P<0.001 vs. siRNA-PTK6 + Ov-NC. (D) TUNEL staining assays detected the apoptosis of C-33A cells. (E) Western blotting was used to assess the levels of Bcl-2, Bax and Birc5 in C-33A cells. **P<0.01 and ***P<0.001 vs. Control or siRNA-PTK6 + Ov-NC. GAB1, GRB2-associated binding 1; PTK6, protein tyrosine kinase 6; RT-qPCR, reverse transcription-quantitative PCR; Ov, overexpression; NC, negative control; siRNA, small interfering RNA; TUNEL, terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling.
Fig 3: Silencing of PTK6 mitigates cervical cancer cell invasion and migration activities. (A) Representative images of wound healing assay (magnification, ×100) and quantification of cell migration. (B) Representative images of Transwell assay (magnification, ×100) and quantification of cell invasion. (C) RT-qPCR and (D) western blotting were used to assess MMP12 and MMP9 expression levels. ***P<0.001 vs. siRNA-NC. PTK6, protein tyrosine kinase 6; RT-qPCR, reverse transcription-quantitative PCR; MMP, matrix metalloproteinase; siRNA, small interfering RNA; NC, negative control.
Fig 4: PTK6 and LRRK2 mediate the role of LINK-A in regulating HIF-1a in RA FLSs.RA FLSs were transfected with LINK-A siRNA (si-LINK-A-2 or si-LINK-A-3) or PTK6 siRNA (si-PTK6-2 or si-PTK6-3) or LRRK2 siRNA (si-LRRK2-1 or si-LRRK2-3) or control siRNA (siC). The mRNA and protein levels were measured using RT-qPCR and Western blot, respectively. (A and B) Effect of LINK-A knockdown with siRNA on the expression of the mRNA (A) and protein (B) of PTK6. (C and D) Effect of LINK-A knockdown on the expression of the mRNA (C) and protein (D) of LRRK2. (E and F) Effect of PTK6 knockdown with siRNA on the expression of the mRNA (E) and protein (F) of HIF-1a. (G and H) Effect of LRRK2 knockdown with siRNA on the expression of the mRNA (G) and protein (H) of HIF-1a. Data (B, D, F, and H, lower or right panel) are expressed as the mean ± SD of densitometry quantification of Western blot from at least 3 independent experiments. *P < 0.05, **P < 0.01, and ***P < 0.001 versus siC, by 1-way ANOVA.
Fig 5: Silencing of PTK6 decreases GAB1 expression. (A) RT-qPCR and (B) western blotting, respectively, determined GAB1 expression in immortalized human normal cervical epithelial cells (Ect1/E6E7) and cervical cancer cell lines (SiHa, HeLa, Ca-Ski, C-33A). (C and D) Co-IP assay was performed using control IgG beads and immunoblotted for PTK6 and GAB1. The expression of GAB1 (E) mRNA and (F) protein levels following PTK6 interference in cervical cancer C-33A cells. ***P<0.001 vs. Ect1/E6E7 or siRNA-NC. PTK6, protein tyrosine kinase 6; GAB1, GRB2-associated binding 1; RT-qPCR, reverse transcription-quantitative PCR; Co-IP, co-immunoprecipitation; siRNA, small interfering RNA; NC, negative control.
Supplier Page from Abcam for Anti-Brk/PTK6 antibody [EPR21051-96]