Experiment Overview
| Repository ID: | FR-FCM-Z6XS | Experiment name: | FACS of SuSa cells with high vs. low MHC-I surface levels | MIFlowCyt score: | 47.00% |
| Primary researcher: | Danielle Hamm | PI/manager: | Danielle Hamm | Uploaded by: | Danielle Hamm |
| Experiment dates: | 2022-01-03 - 2022-01-03 | Dataset uploaded: | Aug 2023 | Last updated: | Aug 2023 |
| Keywords: | [DUX4] [MHC-I] | Manuscripts: | |||
| Organizations: |
Fred Hutchinson Cancer Research Center, Seattle, WA (USA)
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| Purpose: | 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. | ||||
| Comments: | 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. | ||||
| Funding: | Not disclosed | ||||
| Quality control: | None | ||||
