Research Paper Volume 16, Issue 6 pp 5526—5544

Figure 2. USP36 overexpression aggravates malignant phenotypes of colon cancer cells in vitro. (A) The mRNA (top) and protein (button) expression levels of USP36 in HCT116 cells were respectively detected by RT-PCR and western blot to validate the transfection efficiency of USP36 OE. (B, C) The cell proliferation was detected by colony formation (B) and Edu staining (C, Scalebar=100 μm) assays. (D) The cell viability was detected by CCK-8 assay after a series of concentrations of Oxa for the determination of IC50 value to Oxa. (E) The cellular self-renewal capacity was investigated by sphere formation assay (Scalebar = 100 μm). (F) The protein expression of cancer stem cell-related markers (CD133, CD44, Nanog, and Oct-4) was detected by western blot. (G) The cell migration was detected by wound healing assay (Scalebar = 100 μm). (H) The cell invasion was observed by Transwell assay (Scalebar = 100 μm). ^*p < 0.05, ^**p < 0.01, and ^***p < 0.005; Abbreviation: ns: none significance.