FACS of SuSa cells with high vs. low MHC-I surface levels

To determine whether endogenous DUX4 expression in cancer cells correlates with MHC-I suppression. Conclusion: Fluorescence-activated cell sorting paired with RT-qPCR analysis of IFNg-stimulated SuSa cells revealed that cells with low levels of MHC-I had elevated levels of DUX4 target gene expression relative to MHC-I high cells. Notes: SuSa cells were synchronized and released from confluence, lifted using trypsin, seeded onto gelatin-coated plates, treated with or without IFNg for the last 16 hours, and harvested at 72 hours post-release. Cells were stained with BV605 anti-human HLA-A,B,C Antibody (BioLegend #311432), resuspended in FACS buffer (1xDPBS, 1% BSA, 5mM EDTA), and sorted using the BD FACS Aria II with BD FACSDiva software. Data were analyzed using FlowJo V10.5.3.