cck 8 reagent  (MedChemExpress)

Journal: Genes & Diseases

Article Title: Identification of PLXNC1 as a novel biomarker for consensus molecular subtype 4 in colorectal cancer

doi: 10.1016/j.gendis.2025.101974

Figure Lengend Snippet: PLXNC1 promotes the proliferation, migration, and invasion of colorectal cancer (CRC) cells in vitro. (A) qRT-PCR analysis was conducted to examine the knockdown efficiency of PLXNC1 in the negative control (NC)-siRNA- or PLXNC1-siRNA-transfected CRC cells. NC-siRNA was used as a negative control. GAPDH was a normalization control. (B) CCK-8 assay was performed to assay the viability of control and PLXNC1-silenced CRC cells. (C) After 48 h of transfection with NC-siRNA or PLXNC1-siRNA, the migration of CRC cells in Transwell chambers was determined. (D) After 48 h of transfection with NC-siRNA or PLXNC1-siRNA, CRC cells invading the Matrigel in Transwell chambers were performed. Note: Scale bars = 200 μm. (E) Representative images of EdU-stained (red) cells and the quantification of cell proliferation ratio normalized to Hoechst-stained (blue) cells. Scale bars = 75 μm. (F) qRT-PCR data of the PLXNC1 expression in LoVo cells following transfection with PLXNC1 plasmids or controls. GAPDH was a normalization control. (G) CCK-8 assay results show the viability of LoVo cells following transfection with PLXNC1 plasmids or controls. (H, I) Transwell assay was employed to count the number of migrated (H) and invaded (I) LoVo cells following transfection with PLXNC1 plasmids or controls. ∗∗∗ p < 0.001; ∗∗ p < 0.01, and ∗ p < 0.05.

Article Snippet: CCK-8 reagent (10 μL, DOJINDO, Kumamoto, Japan) was added to each well at 0, 24, 48, and 72 h after transfection and incubated at 37 °C for 4 h. Absorbance at 450 nm was measured using a microplate reader (PerkinElmer EnVision, Massachusetts, USA).

Techniques: Migration, In Vitro, Quantitative RT-PCR, Knockdown, Negative Control, Transfection, Control, CCK-8 Assay, Staining, Expressing, Transwell Assay