plasmid flag chk1  (Addgene inc)

Journal: Cell reports

Article Title: ATRX loss in glioma results in dysregulation of cell-cycle phase transition and ATM inhibitor radio-sensitization.

doi: 10.1016/j.celrep.2021.110216

Figure Lengend Snippet: Figure 4. Loss of ATRX in murine neuronal precursor cells (mNPC) and murine GBM cells (mGBM) results in downregulation of Chk1 (A) ATRX promotor binding at Chek1 loci in mNPC cells (ChIP-seq, n = 3, from Danussi et al., 2018). (B) mGBM cells with ATRXKO (‘‘NPA’’) show reduced ATRX and H3.3 binding at Chek1 gene loci (‘‘1’’ through ‘‘4’’ from A) compared to mGBM cell controls without ATRXKO (‘‘NP’’), n = 3. (C) Chek1 expression is downregulated in mNPC cells with ATRX loss. (D) Western blot of U251 ATRXEV and U251 ATRXKO cells with and without 4 Gy IR. (E) Western blot of U251 ATRXKO cells with isogenic Chk1 overexpression or empty vector (n = 3 replicates for C and D). (F and G) Incucyte live-cell imaging analysis of U251 ATRXKOChk1OE cells incorporated with the FastFUCCI reporter plasmid show a gradual return (more than 1.5 times slower) to cycling after 4 Gy IR. (Means ± SEMs for triplicate experiments are shown. *p % 0.05, **p % 0.01, ***p % 0.001, and ****p % 0.0001 using Welch’s t test.) For additional data, see also Figure S4.

Article Snippet: The Chk1 overexpression plasmid was obtained from Addgene (#22894) and was cloned into lentiviral particles at the Vector core at the University of Michigan.

Techniques: Binding Assay, ChIP-sequencing, Expressing, Western Blot, Over Expression, Plasmid Preparation, Live Cell Imaging

Journal: Cell reports

Article Title: ATRX loss in glioma results in dysregulation of cell cycle phase transition and ATM inhibitor radio-sensitization

doi: 10.1016/j.celrep.2021.110216

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: pcDNA4-Chk1-Flag Overexpression Plasmid , Addgene , #22894.

Techniques: Plasmid Preparation, Control, Recombinant, Western Blot, Gene Expression, Inhibition, Generated, Over Expression, Software