In contrast, the gel-based assay directly monitors enzymatic conversion of PARylated PARP1 to an unmodified product by PARG, enabling a semi-quantitative measurement of enzyme inhibition (Supplementary Fig. 4). The IC50 value of a known PARG inhibitor, adenosine diphosphate (hydroxymethyl)pyrrolidine...
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TUNEL assay was also done using the DeadEnd Colorimetric TUNEL System (Promega), according to the manufacturer’s instruction. Images of renal morphology were obtained by light microscopy. Microarray analysis and bioinformatics The integrity of total RNA extracted from the kidney was assessed using a...
Cell viability assay The cell viability analyses were assessed using Annexin V-FITC Apoptosis Detection Kit with PI (Biolegend, San Diego, CA, USA). Briefly, 16–18 h post-UVA irradiation, cells with different pretreatments were washed with PBS and re-suspended in 0.2 ml cold binding buffer....
developed a sequencing technique to analyze the chromatin accessibility of individual cells via assay for transposase-accessible chromatin called as scATAC-seq. They generated DNA accessibility maps from 254 individual GM12878 lymphoblastoid cells and discovered combinations of trans-factors associated with ...
Classically, cell death is categorized into three different types, based on morphological changes, triggers, and the biochemical pathways involved. Apoptosis, or Type I cell death, is a tightly regulated form of programmed cell death (PCD), that triggers cells to self-destruct without any ...
Luciferase reporter assay Before transfection, 1 × 105 GSCs were seeded in a 24-well dish. The following day, cells were co-transfected with the control pcDNA3.1 empty vector (pcDNA3.1, 225 ng) or ATF3 plasmid (225 ng)-based firefly luciferase reporter and the Renilla luciferase ...
such as Parp1 inhibitors, and that Cdk12 inhibitors may induce sensitization of HR-competent tumors to these treatments.6,7,10,11 Despite growing clinical interest, the mechanism by which Cdk12 regulates HR genes remains unfamiliar. Here we find that Cdk12 globally suppresses intronic polyadenylatio...
Here, we adapted the Multicolor Competition Assay (MCA)23, based on human cells depleted in each of the main DNA repair pathways (Table 1), to study the repair mechanisms involved in the cellular response to genotoxic treatments. The implication of each DNA repair mechanism involved in CDT res...
Live-cell-imaging-based screens identify RasGEFX as an essential component of the pathway that triggers spontaneous Ras excitation for basal cell migration in combination with RasGEFB/M/U and for macropinocytic cup formation and fluid uptake. ...