Journal: Molecular cell

Article Title: Targeted and Persistent 8-Oxoguanine Base Damage at Telomeres Promotes Telomere Loss and Crisis

doi: 10.1016/j.molcel.2019.04.024

Figure Lengend Snippet: (A) Endogenous OGG1 and OGG1-EGFP (red) localization to telomeres (green) in HeLaFAP cells by anti-OGG1 IF and telo-FISH after dye + light.

Article Snippet: After resuspension in TE, DNA was treated with FPG (NEB, 2.7U/ μg DNA), OGG1 (Novus, 125 nM), and APE1 (NEB, 3.3U/ μg DNA) enzymes alone or in combination for 2 hours at 37°C in 1X Cutsmart buffer (NEB).

Techniques:

Journal: Molecular cell

Article Title: Targeted and Persistent 8-Oxoguanine Base Damage at Telomeres Promotes Telomere Loss and Crisis

doi: 10.1016/j.molcel.2019.04.024

Figure Lengend Snippet: (A) Immunoblot of OGG1 in parental HeLaFAP cells and two OGG1 KO clones. Actin was used as a loading control.

Article Snippet: After resuspension in TE, DNA was treated with FPG (NEB, 2.7U/ μg DNA), OGG1 (Novus, 125 nM), and APE1 (NEB, 3.3U/ μg DNA) enzymes alone or in combination for 2 hours at 37°C in 1X Cutsmart buffer (NEB).

Techniques: Western Blot, Clone Assay, Control

Journal: Molecular cell

Article Title: Targeted and Persistent 8-Oxoguanine Base Damage at Telomeres Promotes Telomere Loss and Crisis

doi: 10.1016/j.molcel.2019.04.024

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: After resuspension in TE, DNA was treated with FPG (NEB, 2.7U/ μg DNA), OGG1 (Novus, 125 nM), and APE1 (NEB, 3.3U/ μg DNA) enzymes alone or in combination for 2 hours at 37°C in 1X Cutsmart buffer (NEB).

Techniques: Recombinant, Plasmid Preparation, Imaging, Clone Assay, Sequencing, shRNA, Software

Journal: Scientific Reports

Article Title: Metabolic signatures of germination triggered by kinetin in Medicago truncatula

doi: 10.1038/s41598-019-46866-6

Figure Lengend Snippet: ( a ) Alkaline comet assay was used to measure the accumulation of DNA strand breaks in M . truncatula seeds collected at the radicle protrusion stage. ( b ) Transcript levels of MtOGG1 and MtFPG genes in seeds collected at the radicle protrusion stage determined by q RT-PCR analyses. ( c ) EPR spectrum generated by kinetin-treated M . truncatula seeds collected at the radicle protrusion stage. The enlargement of the central part of the same EPR spectrum shows those regions corresponding to reactive oxygen species (ROS) and organic radicals, respectively. The EPR spectrum was recorded at low field. For the iron region (range from 100 to 550 mT) an applied microwave power of 10 mW with a modulation amplitude of 0.5 mT was used. For organic radicals and Mn 2+ , a range from 331 to 341 mT and an applied microwave power of 10 mW and a modulation amplitude of 0.2 mT were applied. ( d ) Transcript levels of MtAPX , MtMT2 , MtTRH1 and MtTRH2 genes in seeds collected at the radicle protrusion stage determined by q RT-PCR analyses.Values are expressed as mean ± SD of three independent replications with 20 seeds for each replication. Asterisks indicate statistically significant differences determined using Student’s t -test ( P < 0.05). a.u., arbitrary units. W, control seeds imbibed with water. Kin, seeds imbibed with 0.5 mM kinetin. APX, ascorbate peroxidase. FPG, formamidopyrimidine-DNA glycosylase. MT2, type 2 metallothionein; OGG1, 8-oxoguanine DNA glycosylase.TRH, thioredoxin. mT, milliTesla.

Article Snippet: Studies carried in the model legume Medicago truncatula have demonstrated that the DNA damage response triggered during early seed imbibition features the up-regulation of key BER (base excision repair) genes, as OGG1 (8-oxoguanine glycosylase/lyase) and FPG (formamidopyrimidine DNA glycosylase) required to remove the DNA oxidative lesions under physiological and stress conditions , , .

Techniques: Alkaline Single Cell Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Generated, Control