Figure 3
From: Olig2 regulates p53-mediated apoptosis, migration and invasion of melanoma cells
Downregulation of Olig2 induced p53-mediated apoptosis. (a) Apoptotic cells were detected using RealTime Glo Annexin V apoptosis assay kit. 24 h after transfection, the intensity of luminescence was recorded using a luminescence plate reader (**p < 0.01; ***p < 0.001 versus Con cells, n = 3). (b) Protein expression of Olig2, p53 and Bcl-2 was detected in A375 and 501mel melanoma cells by western blot analysis with Olig2 (H-10) antibody. β-actin was used as loading control. The band intensity was quantified using NIH ImageJ software 1.45 s. (c) Protein expression of Olig2 and p21 in A375 and 501mel melanoma cells by western blot analysis. (d) The activities of caspases-3/-7 were evaluated using Caspase-Glo 3/7 assay kit. 24 h after transfection, fluorescence intensity was detected at 485 nm excitation and 520 nm emission on a fluorescence plate reader (***p < 0.001 versus Con cells, n = 3). (e) PARP and cleaved PARP expression in two melanoma cells transfected with negative control (Con) or CRISPR/Cas9-Olig2 plasmid (CRISPR-Olig2) (1 μg and 2 μg, respectively) was detected by western blot analysis. Original images of blots were provided in supplementary file.
