Circadian Imaging of PER and CRY Proteins in U-2 OS cells

ABSTRACT: U-2 OS knock-in cells were genereated using CRISPR to express CRY1, CRY2, PER1 PER2 fused to either mScarlet-I or mClover3 from the endogenous promoter. All cells were transduced with H2B-iRFP using lentivirus to allow segmentation and tracking of nuclei. Additionally, cells were transduced with lentivirus expressing shRNA against either FBXL3 (pGIPZ V2LHS_254986) or a nontargeted control shRNA. ===Raw Data=== ==Dataset Dex== Cells of selected clones were imaged for ~3days after synchronization with dexamethasone (1µM, 20min) at an imaging interval of 1 picture per hour ==Dataset CHX/Main== In 3 experiments, cells were imaged for 68 hours with an imaging interval of 1 picture per hour. After 44-48 hours, 20 µg/ml CHX was added to a subset of cells to inhibit protein production. For as

ORGANISM(S): Homo sapiens (human)

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PROVIDER: S-BIAD1038 | bioimages |

REPOSITORIES: bioimages