Figure 3. High-content screening (HCS) of YC-1-treated cells under chemical hypoxia. 661W cells were exposed to 0.066% DMSO, 20 μM YC-1, 200 μM CoCl₂, or both YC-1 and CoCl₂ for 24 h. After fixation and immunofluorescence staining, cells were detected and analyzed using a Cellomics ArrayScan VTI HCS Reader. A: Representative fluorescent photographs show the distribution of K_i-67 (green) and Hoechst-labeled nuclei (blue) using the Columbus Image Data Storage and Analysis System. Panels (B) and (C) show quantified HCS results for the number and ratio of K_i-67-stained nuclei (n=4). * Indicates p<0.05 compared to the control group or the DMSO vehicle group; # indicates p<0.05 compared to the chemically induced hypoxic group.