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Image Search Results
Journal: bioRxiv
Article Title: Elongation factor ELOF1 drives transcription-coupled repair and prevents genome instability
doi: 10.1101/2021.05.11.443558
Figure Lengend Snippet: (A) Left panel: Schematic of the genomic locus of CSB and used strategy for generating the homozygous CSB-mScarletI-HA KI cell line. Half arrows indicate primer locations. Middle and right panel: Genotyping PCR and immunoblot for CSB -KI cell line. (B) Left panel: Schematic of the genomic locus of UVSSA and used strategy for generating the homozygous UVSSA-mScarletI-HA KI cell line. Half arrows indicate primer locations. Middle and right panel: Genotyping PCR and immunoblot for UVSSA -KI cell line. (C) Left panel: CSB mobility was determined by FRAP analysis of CSB-mScarletI after the indicated treatments. THZ1: 1 hour treatment (2 µM) before UV-C irradiation (4 J/m 2 ) or mock treatment. Right panel: Relative immobile fraction of CSB as determined by FRAP analysis. Plotted values represent mean ± SEM and are normalized to mock treated. n≥15 cells. (D) Same as C but for UVSSA-mScarletI. n≥10 cells. (E+F) FRAP analyses of CSB-mScarletI (E) or UVSSA-mScarletI (F) mobility after transfection with indicated siRNAs in individual graphs. Cells were mock treated (NT) or analyzed directly (UV) or 5 hours (5hr UV) after irradiation with 4 J/m 2 UV-C. (G) Relative fluorescence intensity of UVSSA in UVSSA -KI cells transfected with indicated siRNAs as determined by live-cell imaging. Plotted values represent mean ± SEM. n≥16 cells. (H) FRAP analysis of CSB in CSB -KI cells transfected with indicated siRNAs 2 hours after UV. VCPi: VCP inhibitor (5 µM) was directly added after UV-C (4 J/m 2 ). (I) Immunoblot of chromatin fraction of indicated cell lines 1 hour after 12 J/m 2 UV-C or mock treatment. NAEi = 1 hour treatment with NEDDylation inhibitor (10 µM). SSRP1 is shown as loading control. (J) Immunoblot of chromatin fraction of HCT116 cells transfected with indicated siRNAs 1 hour after 12 J/m 2 UV-C or mock treatment. SSRP1 is shown as loading control.
Article Snippet: Cell were treated for 1 hour with the following chemicals: Actinomycin D (Sigma, 1 µg/ml), Flavopiridol (Sigma, 1 µM),
Techniques: Western Blot, Irradiation, Transfection, Fluorescence, Live Cell Imaging