Journal: Journal of Clinical Laboratory Analysis

Article Title: Upregulation of KIF18B facilitates malignant phenotype of esophageal squamous cell carcinoma by activating CDCA8 / mTORC1 pathway

doi: 10.1002/jcla.24633

Figure Lengend Snippet: The effects of KIF18B overexpression on EMT‐related proteins and mTORC1 pathway‐related proteins in ESCC cells. Both EC9706 and KYSE220 cells were transfected with KIF18B overexpression plasmid. (A–D) Western blot was employed to examine the expression of EMT biomarkers, including E‐cadherin, N‐cadherin and Vimentin. GAPDH served as a loading control. (E–K) The levels of CDCA8 and mTORC1 pathway‐related proteins (p‐p70S6K, p70S6K, p‐mTOR, and mTOR) were detected by western blot. GAPDH served as a loading control. All experiments had gone through three repetitions. ** p < .01, *** p < .001 versus Vector. EMT, epithelial mesenchymal transition; ESCC, esophageal squamous cell carcinoma; GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase; KIF18B, kinesin family member 18B; mTORC1, mechanistic target of rapamycin complex 1; p, phosphorylated

Article Snippet: The primary antibodies adopted here encompassed those against KIF18B (ab168812, 94 kDa, Abcam, UK, 1:2000), CDK4 (ab108357, 34 kDa, Abcam, UK, 1:1000), CyclinD3 (ab28283, 35 kDa, Abcam, UK, 1:1000), p70S6K (ab32529, 70 kDa, Abcam, UK, 1:5000), phosphorylated (p)‐p70S6K (ab59208, 60 kDa, Abcam, UK, 1:1000), p‐mTOR (ab109268, 289 kDa, Abcam, UK, 1:1000), mTOR (ab134903, 289 kDa, Abcam, UK, 1:10,000), E‐Cadherin (ab40772, 97 kDa, Abcam, UK, 1:10000), Vimentin (ab92547, 54 kDa, Abcam, UK, 1:1000), N‐Cadherin (ab18203, 130 kDa, Abcam, UK, 1:1000), CDCA8 (ab74473, 38 kDa, Abcam, UK, 1:1000), and GAPDH (ab8245, 36 kDa, Abcam, UK, 1:2000).

Techniques: Over Expression, Transfection, Plasmid Preparation, Western Blot, Expressing, Control