Journal: Communications Biology

Article Title: Three-dimensional CRISPR screening reveals epigenetic interaction with anti-angiogenic therapy

doi: 10.1038/s42003-021-02397-3

Figure Lengend Snippet: a Effects of NSC 663284 or TMP195 on survival and/or proliferation of ECs treated with JQ1 plus bevacizumab or palivizumab in ESFM. The addition of NSC 663284 at 10 µM further reduced viable cell number in ECs treated with JQ1 plus bevacizumab while the addition of TMP195 did not affect those treated with JQ1 plus palivizumab. b Relative cell response to bevacizumab in ESFM plus DMSO, NSC 663284, or TMP195. The addition of NSC 663284 or TMP195 did not affect EC response to bevacizumab. Error bars represent ± SEM ( n = 2 independent experiments). **adjusted P value < 0.01; ns, not significant (versus B + JQ1, P + JQ1 or DMSO, one-way ANOVA with Dunnett’s multiple comparison test performed). c Schematic model illustrating a potential role for CDC25B in regulating the EC response to co-treatment with BETi plus bevacizumab. Top (palivizumab plus BETi), low CDC25B expression results in phosphorylated cyclin-dependent kinase (CDK) 1/2 and hyperactive centromere protein (CENP)-A deposition, which can cause centromere defects and chromosomal instability. Middle (bevacizumab plus BETi), upregulated CDC25B expression caused by co-treatment dephosphorylates and activates CDK1/2, which reduces CENP-A deposition by phosphorylating the M18BP1 and Holiday junction recognition protein (HJURP) complexes. Eventually, centromere function and chromosomal stability could be improved. Bottom (bevacizumab plus BETi plus NSC 663284), inhibition of CDC25B leads to low CDK1/2 activities in the presence of bevacizumab plus BETi. Reduced CENP-A deposition is therefore restored, which could result in centromere defects and chromosomal instability.

Article Snippet: The pan-BETi JQ1 and I-BET762 (Selleckchem or MedChemExpress), the small-molecule TLK1 inhibitor thioridazine HCl (THD; Sigma), the small-molecule CDC25 dual specificity phosphatase inhibitor NSC 663284 (MedChemExpress), and the small-molecule selective Class IIa HDAC inhibitor TMP195 (MedChemExpress) was reconstituted and stored according to the manufacturer’s instructions.

Techniques: Comparison, Expressing, Inhibition

Journal: International Journal of Medical Sciences

Article Title: Cell Recognition Molecule L1 Regulates Cell Surface Glycosylation to Modulate Cell Survival and Migration

doi: 10.7150/ijms.20479

Figure Lengend Snippet: Effects of inhibitors of signal transduction pathways on cell migration, cell survival and the protein expressions of FUT9 and ST6Gal1. After incubation for 1 hour of L1-CHO cells treated with or without L1Ab, inhibitors of PLCγ (U73122, 10.5 μM), CDC25 phosphatase (CDC25 inhibitor II, 1.05 μM), PI3K (LY294002, 16.5 μM), Erk (Erk inhibitor, 50 μM), JNK (JNK inhibitor, 200nM), ATM/ATR (Caffeine, 1mM) or PKA inhibitor (KT5720, 280 nM) were added into the culture medium and the cells were incubated further for 24 hours. A. After treatment with inhibitors of the signal transduction pathways, cell migration assay was performed. Cell migration was significantly inhibited in L1Ab-treated L1-CHO cells by LY294002 and Erk inhibitor. B. After treatment with inhibitors of the signal transduction pathways, cell survival was quantified by MTT assay. U73122, LY294002, JNK inhibitor, Caffeine and Erk inhibitor significantly repressed cell survival in L1-CHO cells. C. Western blot was used to detect the expression of transferases. The protein expressions of ST6Gal1 and FUT9 were significantly downregulated in L1-CHO cells treated with LY294002 and Erk inhibitor. * : p <0.05; ** : p <0.01, by Student's test comparison with the no inhibitor control.

Article Snippet: Co. Ltd., Japan), Tunicamycin (Calbiochem, CA, USA), U73122 (Calbiochem, CA, USA), Cdc25 phosphatase inhibitor II (Calbiochem, CA, USA), LY294002 (Sigma, MO, USA), Erk inhibitor (3-(2-Aminoethyl)-5-((4-ethoxyphenyl) methylene) -2,4-thiazolidinedione, HCl; Calbiochem, CA, USA) and KT5720 (Sigma, MO, USA) are available commercially.

Techniques: Transduction, Migration, Incubation, Cell Migration Assay, MTT Assay, Western Blot, Expressing

Journal: Frontiers in Genetics

Article Title: WIP1 Contributes to the Adaptation of Fanconi Anemia Cells to DNA Damage as Determined by the Regulatory Network of the Fanconi Anemia and Checkpoint Recovery Pathways

doi: 10.3389/fgene.2019.00411

Figure Lengend Snippet: Boolean functions for the nodes in the FA-CHKREC network.

Article Snippet: We tested this model-derived hypothesis in the EUFA316+EV cell line, a lymphoblast cell line derived from a FA patient with inherited mutations in FANCG , using chemical inhibitors against the components of the CHKREC nodes, namely GSK830371 (Sigma-Aldrich) for WIP1 inhibition, TC-S 7010 (Selleckchem) for inhibiting Aurora A, BI2536 (Selleckchem) for inhibiting PLK1 and CDC25 Phosphatase Inhibitor II (Santa Cruz Biotechnologies) for inhibiting CDC25. shows survival curves from the different CHKREC inhibitors tested.

Techniques:

Journal: Frontiers in Genetics

Article Title: WIP1 Contributes to the Adaptation of Fanconi Anemia Cells to DNA Damage as Determined by the Regulatory Network of the Fanconi Anemia and Checkpoint Recovery Pathways

doi: 10.3389/fgene.2019.00411

Figure Lengend Snippet: Biological meaning of the attractors obtained in the FA-CHKREC network.

Article Snippet: We tested this model-derived hypothesis in the EUFA316+EV cell line, a lymphoblast cell line derived from a FA patient with inherited mutations in FANCG , using chemical inhibitors against the components of the CHKREC nodes, namely GSK830371 (Sigma-Aldrich) for WIP1 inhibition, TC-S 7010 (Selleckchem) for inhibiting Aurora A, BI2536 (Selleckchem) for inhibiting PLK1 and CDC25 Phosphatase Inhibitor II (Santa Cruz Biotechnologies) for inhibiting CDC25. shows survival curves from the different CHKREC inhibitors tested.

Techniques: Activation Assay

Journal: Frontiers in Genetics

Article Title: WIP1 Contributes to the Adaptation of Fanconi Anemia Cells to DNA Damage as Determined by the Regulatory Network of the Fanconi Anemia and Checkpoint Recovery Pathways

doi: 10.3389/fgene.2019.00411

Figure Lengend Snippet: Inactivation of CHKREC nodes in FA mutants promotes CCA and reduces FA cell survival. (A) Double KO simulations of the FAcore and components of the CHKREC (WIP1, CDK1-AurA, PLK1, CDC25, and CycB-CDK1) showing that FA cell division will be blocked since the CycB-CDK1 node cannot be activated, driving the system to a cyclic CCA attractors. Only attractors are shown. Nodes in the simulations are grouped by color according to functional categories: DNA damage in black, DNA repair pathways in blue, Checkpoint in red and CHKREC in green. Inactive nodes are colorless, whereas active nodes are colored according to their functional category. (B) Schematics showing that upon CHKREC inhibition, the division of FA mutant cells with unrepaired DNA damage will be blocked and the cell will remain in a CCA attractor. In biological terms, cell division blockade may drive the cell to senescence or cell dead. (C) Screening of multiple CHKREC chemical inhibitors showing that the FAcore mutant cell line EUFA316+EV ( FANCG deficient) is more sensitive to CHKREC inhibition than its corrected counterpart EUFA316+G. Refer to , , to see the trajectories followed by these and other FAcore and FANCD2I double null mutants, respectively, before arriving to an attractor.

Article Snippet: We tested this model-derived hypothesis in the EUFA316+EV cell line, a lymphoblast cell line derived from a FA patient with inherited mutations in FANCG , using chemical inhibitors against the components of the CHKREC nodes, namely GSK830371 (Sigma-Aldrich) for WIP1 inhibition, TC-S 7010 (Selleckchem) for inhibiting Aurora A, BI2536 (Selleckchem) for inhibiting PLK1 and CDC25 Phosphatase Inhibitor II (Santa Cruz Biotechnologies) for inhibiting CDC25. shows survival curves from the different CHKREC inhibitors tested.

Techniques: Functional Assay, Inhibition, Mutagenesis