Journal: Cancer Cell

Article Title: A Dual Role of Caspase-8 in Triggering and Sensing Proliferation-Associated DNA Damage, a Key Determinant of Liver Cancer Development

doi: 10.1016/j.ccell.2017.08.010

Figure Lengend Snippet:

Article Snippet: p-Chk2 [p Thr68] Rabbit polyclonal Antibody , Novus Biologicals , Cat# NB100-92502; RRID: AB_1216474.

Techniques: Purification, Imaging, Virus, Control, Mutagenesis, Recombinant, Staining, Reverse Transcription, SYBR Green Assay, Microarray, RNA Expression, RNA Sequencing, Methylation, Labeling, Software, Light Microscopy

Journal: Oncology reports

Article Title: Novel SAHA‑bendamustine hybrid NL‑101 in combination with daunorubicin synergistically suppresses acute myeloid leukemia.

doi: 10.3892/or.2020.7591

Figure Lengend Snippet: Figure 3. NL‑101 in combination with DNR promotes DNA damage‑related signaling. MV4‑11 cells were treated with 300 nM NL‑101, 8 nM DNR and the combination; HL‑60 cells were treated with 500 nM NL‑101, 16 nM DNR and the combination for 48 h. Western blot analysis was conducted for γ‑H2AX, ATR, p‑ATR, ATM, p‑ATM, CHK1, p‑CHK1, CHK2 and p‑CHK2 protein levels. DNR, daunorubicin; p, phosphorylated.

Article Snippet: Antibodies against caspase-3 (cat. no. 9662), caspase-7 (cat. no. 9492), PARP (cat. no. 9532), BAD (cat. no. 9239), BIM (cat. no. 2933), cyclin B1 (cat. no. 4135), cell division cycle protein 2 (CDC2; cat. no. 9116), γ-H2AX (Ser139; cat. no. 9718), phosphorylated (p)-ATR (Thr1981; cat. no. 30632), p-ATM (Ser1981; cat. no. 5883), p-CHK1 (Ser345; cat. no. 2348), p-CHK2 (Thr68; cat. no. 2197), GAPDH (cat. no. 5174) and β-actin (cat. no. 4970) were purchased from Cell Signaling Technology, Inc. ATR (cat. no. 19787-1-AP), ATM (cat. no. 27156-1-AP), CHK1 (cat. no. 10362-1-AP) and CHK2 (cat. no. 13954-1-AP) antibodies were purchased from ProteinTech Group, Inc. Anti-human (h) CD45‐fluorescein isothiocyanate (FITC) (cat. no. ab134199) was obtained from Abcam.

Techniques: Western Blot

Journal: Endocrinology

Article Title: The role of exchange protein directly activated by cyclic AMP 2-mediated calreticulin expression in the decidualization of human endometrial stromal cells.

doi: 10.1210/en.2013-1478

Figure Lengend Snippet: Figure 4. Knock-down of EPAC2 or CRT in ESCs is associated with senescence. ESCs were treated for 24 hours with nontargeting control (Cont), EPAC2, or CRT siRNA and then cultured for 72 hours. A, B, ESCs were stained with SA--Gal (A). Scale bars, 100 m. The graph shows the relative levels of SA--Gal-positive cells. The data from three independent experiments are presented as ratios and show the mean SEM (B). *, P .01 vs Cont. C, D, Cell lysates were subjected to immunoblot analysis with anti-p53, p21, CRT, or EPAC2 antibody. The same blot was stripped and reprobed with anti-GAPDH antibody as a loading control (C). The graphs show the relative levels of p53 or p21 normalized to GAPDH levels (D). The data (mean SEM) from three independent experiments are presented as ratios. *, P .01 vs Cont.

Article Snippet: Antibodies against CRT, p53 (clone 1C12), p21 (clone DCS60), EPAC1 (clone 5D3), and EPAC2 (clone 5B1) were purchased from Cell Signaling Technology.

Techniques: Knockdown, Control, Cell Culture, Staining, Western Blot

Journal: Biological & pharmaceutical bulletin

Article Title: Glionitrin A, a new diketopiperazine disulfide, activates ATM-ATR-Chk1/2 via 53BP1 phosphorylation in DU145 cells and shows antitumor effect in xenograft model.

doi: 10.1248/bpb.b13-00719

Figure Lengend Snippet: Fig. 2. Glionitrin A Elevates H2A.X Phosphorylation by Responding to DNA Damage Resulting from the Activation of ATM/ATR-Chk1/2 in a 53BP1-Dependent Manner

Article Snippet: Primary antibodies against phospho-H2A.X (Ser139), phospho-53BP1 (S11778), 53BP1, phospho-ATM (Ser1981), phospho-ATR (Ser428), ATR, phospho-Chk1 (Ser317), phospho-Chk1 (Ser345), Chk1, phospho-Chk2 (Thr68), phosphoChk2 (Thr387), Chk2, cdc25A, phosphor-cdc2 (Tyr15), cdc2, cleaved caspase-8, Bid, Bax, cleaved caspase-9, cleaved caspase-3, poly(ADP-ribose) polymerase (PARP), apoptosisinducing factor (AIF), endonuclease G (EndoG), histone deacetylase 1 (HDAC1), cytochrome c oxidase subunit IV (COX IV) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and peroxidase-conjugated secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, U.S.A.).

Techniques: Phospho-proteomics, Activation Assay

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Effect of circadian clock mutations on DNA damage response in mammalian cells.

doi: 10.4161/cc.21771

Figure Lengend Snippet: Figure 8. PER1 expression neither sensitizes human lung cancer cells to apoptosis nor affects Chk2 phosphorylation following ionizing radiation (IR). siRNA transfected NCI-H460 human lung cancer cells were irradiated with 0, 10 or 30 Gy of IR and allowed to grow for 48 h, protein lysates were prepared for immunoblot analysis. (A) Protein levels were analyzed by immunoblotting 48 h after irradiation with IR with the indicated antibodies from NCI-H460 cells transfected with either negative control (non-target, NT) or Per1 siRNAs. (B) Quantitative analysis of the immunoblots. (C) Protein levels were analyzed by immunoblotting 48 h after irradiation with IR with the indicated antibodies from NCI-H460 cells transfected with either empty vector (pCDNA3) or a mPer1-overexpressing vector for 72 h. (D) Quantitative analysis of the immunoblots. Averages and standard deviations are from at least three independent experiments. T-PARP and C-PARP stand for total PARP and cleaved PARP, respectively.

Article Snippet: Protein lysates from whole-cell lysates were prepared as described previously,42 and the protein expression levels were determined by immunoblot assay.42 The following antibodies were used to detect the respective proteins: XPA, Cyclophilin-B, p53, Chk1, Chk2 and actin (Santa Cruz Biotechnology); cleaved PARP, cleaved caspase-3, P-chk1, P-chk2 and GAPDH (Cell Signaling Technology); Cry2, Per2, Clock and Bmal1 (Bethyl Labs); Per1 (Kerafast) and Cry1.38 IR dose (0, 2, 2.5, 4, 5, 6, 7.5, 8 and 10 Gy).

Techniques: Expressing, Phospho-proteomics, Transfection, Irradiation, Western Blot, Negative Control, Plasmid Preparation