Fig 1: Effect of NRSN2 overexpression on the migration and invasion of SW620 cells. (A) Wound-healing assay was performed to (B) analyze the migratory ability of SW620 cells transfected with Ov-NRSN2 (magnification, ×100). (C) Transwell assay was employed to (D) analyze the invasive ability of SW620 cells transfected with Ov-NRSN2 (magnification, ×100). (E) MMP2 and MMP protein expression levels were analyzed via western blotting. Data are presented as the mean ± SEM from three independent experiments. **P<0.01 and ***P<0.001 vs. control; ##P<0.01, ###P<0.001 vs. Ov-NC. Ov, overexpression; NRSN2, neurensin-2; NC, negative control.
Fig 2: Effect of NRSN2 overexpression on the proliferation of SW620 cells. (A) Reverse transcription-quantitative PCR and (B) western blotting were performed to evaluate the mRNA and protein expression levels of NRSN2 to determine transfection efficiency of Ov-NRSN2, respectively. (C) Cell Counting Kit-8 assay was used to detect the viability of SW620 cells transfected with Ov-NRSN2. (D) Colony formation assay was employed to determine proliferation ability of SW620 cells transfected with Ov-NRSN2 and (E) colony formation ability from a single cell captured with an inverted fluorescence microscope (magnification, ×400). Data are presented as the mean ± SEM from three independent experiments. **P<0.01 and ***P<0.001 vs. control; ##P<0.01, ###P<0.001 vs. Ov-NC. Ov, overexpression; NRSN2, neurensin-2; NC, negative control; OD, optical density.
Fig 3: NRSN2 is highly expressed in CRC cells and NRSN2-knockdown inhibits the proliferation of SW620 cells. (A) mRNA expression of NRSN2 in CRC cells was evaluated via RT-qPCR. (B) mRNA expression of NRSN2 was evaluated via RT-qPCR to determine the transfection efficiency of shRNA-NRSN2. (C) Cell Counting Kit-8 assay was used to detect the viability of SW620 cells transfected with shRNA-NRSN2. (D) Correlation analysis was used to analyze the correlation between NRSN2 expression and cell viability in SW620 cells. (E) Colony formation assay was performed to determine the proliferative ability of SW620 cells transfected with shRNA-NRSN2 and (F) colony formation ability from a single cell captured with an inverted fluorescence microscope (magnification, ×400). Data are presented as the mean ± SEM from three independent experiments. **P<0.01 and ***P<0.001 vs. FHC or control; ###P<0.001 vs. shRNA-NC. RT-qPCR, reverse transcription-quantitative PCR; shRNA, short hairpin RNA; NRSN2, neurensin-2; OD, optical density; NC, negative control.
Fig 4: Association between NRSN2 and SOX12 in SW620 cells. (A) Interaction between NRSN2 and SOX12 was predicted using the STRING database. (B) IP was performed to confirm the association between NRSN2 and SOX12. (C) SOX12 protein expression was analyzed via western blotting. Data are presented as the mean ± SEM from three independent experiments. ***P<0.001 vs control; ###P<0.001 vs. shRNA-NC. @@@P<0.001 vs. Ov-NC. IP, immunoprecipitation; Ov, overexpression; NRSN2, neurensin-2; NC, negative control; shRNA, short hairpin RNA.
Fig 5: NRSN2 knockdown suppresses and NRSN2 overexpression increases the proliferation of breast cancer cells in vitro. (A) NRSN2 knockdown decreased NRSN2 expression in MDA-MB-231 cells. (B) NRSN2 overexpression increased NRSN2 expression in MDA-MB-231 cells. (C) NRSN2 knockdown inhibited MDA-MB-231 cell proliferation. (D) NRSN2 overexpression promoted MDA-MB-231 cell proliferation. (E) NRSN2 knockdown inhibited the colony formation of MDA-MB-231 cells. (F) NRSN2 overexpression promoted the colony formation of MDA-MB-231 cells. NC, negative control; NRSN2, neurensin-2; ns, not significant; si, small interfering (RNA); siR-NRSN2, siRNA against NRSN2; pRK5-NRSN2, NRSN2 overexpression vector. **P<0.01.
Supplier Page from Abcam for Anti-NRSN2 antibody