other:Article Title: DNA damaging agents boost the transcription of endothelin A receptor in high-grade serous ovarian cancer
Article Snippet: The following primary ab were adopted: anti-ET A R (1:5000, cat. no. PA3-065, Invitrogen); anti-γH2A.X (phospho-gamma-H2A.X Ser-139) (1:1000, cat. no. 2577, Cell Signalling); anti-pATR (Ser428) (1:1000, cat. no. 2853, Cell Signalling); anti-ATR (1:1000; cat. no. 2790; Cell Signalling); anti-pATM (Ser1981) (D25E5) (1:1000, cat. no. 13050, Cell Signalling); anti-ATM (D2E2) (1:1000, cat. no. 2873, Cell Signalling); anti-pCHK1 (Ser345) (133D3) (1:1000, cat. no. 2348, Cell Signalling); anti-CHK1 (2G1D5) (1:1000, cat. no. 2360, Cell Signalling); anti-pCHK2 (T68) (C13C1) (1:1000, cat. no. 2197, Cell Signalling); anti-CHK2 (1:1000, cat. no. 2662, Cell Signalling); anti-pSTAT3 (Y705) (D3A7) (1:1000; cat. no. 9145, Cell Signalling); anti-STAT3 (D3Z2G) (1:1000, cat. no. 12640, Cell Signalling); anti-caspase 3 (D3R6Y) (1:1000, cat. no. 142200, Cell Signalling); anti-cleaved-caspase 3 (D175) (1:1000, cat. no. 9661, Cell Signalling); anti-cleaved-PARP (Asp 214) (1:1000, cat. no. 9541, Cell Signalling); anti-tubulin (DM1A) (1:200, cat. no. sc-32293, Santa Cruz Biotechnology) and anti-β-actin (AC-15) (1:5000, cat. no. 1978, Sigma-Aldrich).
Article Title: NPM1 phosphorylation-mediated telomere maintenance via stabilization of POLD3 in ALT-positive osteosarcoma: unraveling mechanisms and therapeutic opportunities
Article Snippet: Antibodies used in this study were as follows: anti-NPM1 (YT3208 Immunoway, 1:1000 and Cat No. 60096-1-lg proteintech, 1:500), anti-STN1 (ab252855 Abcam, 1:500), anti-γ-H2AX (05-636 Milipore, 1:1000), anti-53BP1 (ab175933 Abcam, 1:1000), anti-ATM (2873S Cell Signaling Technology, 1:1000), anti-ATR (2790S Cell Signaling Technology, 1:1000), anti-Chk1 (ab47574 Abcam, 1:500), anti-p-ATM (5883S Cell Signaling Technology, 1:1000), anti-p-ATR (2853S Cell Signaling Technology, 1:1000), anti-p-Chk1 (12302S Cell Signaling Technology, 1:1000), anti-pT199-NPM1 (3541S Cell Signaling Technology, 1:1000), anti-POLD3 (21935-1-AP Proteintech 1:1000), anti-RAD52 (sc-365341 Santa Cruz, 1:100), anti-GAPDH (AF0911 Affinity, 1:5000), anti-β-tubulin (AC008 Abclonal, 1:2000), anti-HA-tag (3724S Cell Signaling Technology, 1:1000), anti-Flag-tag (2368S Cell Signaling Technology, 1:500), anti-GST-tag (10000-0-AP Proteintech, 1:1000), anti-His-tag (AH367 Beyotime, 1:1000), anti-PML (sc-5621 Santa Cruz, 1:500), Alexa Fluor 555 goat anti-rabbit (A21428 Lifetechnologies, 1:2000), Alexa Fluor 555 goat anti-mouse (A21422 Lifetechnologies, 1:2000), Alexa Fluor 488 goat anti-rabbit (A11008 Lifetechnologies, 1:2000), Alexa Fluor 4488 goat anti-mouse (A11001 Lifetechnologies, 1:2000).
Article Title: DNA damaging agents boost the transcription of endothelin A receptor in high-grade serous ovarian cancer.
Article Snippet: The following primary ab were adopted: anti-ETAR (1:5000, cat. no. PA3-065, Invitrogen); anti-γH2A.X (phospho-gammaH2A.X Ser-139) (1:1000, cat. no. 2577, Cell Signalling); anti-pATR (Ser428) (1:1000, cat. no. 2853, Cell Signalling); anti-ATR (1:1000; cat. no. 2790; Cell Signalling); anti-pATM (Ser1981) (D25E5) (1:1000, cat. no. 13050, Cell Signalling); anti-ATM (D2E2) (1:1000, cat. no. 2873, Cell Signalling); anti-pCHK1 (Ser345) (133D3) (1:1000, cat. no. 2348, Cell Signalling); anti-CHK1 (2G1D5) (1:1000, cat. no. 2360, Cell Signalling); anti-pCHK2 (T68) (C13C1) (1:1000, cat. no. 2197, Cell Signalling); anti-CHK2 (1:1000, cat. no. 2662, Cell Signalling); anti-pSTAT3 (Y705) (D3A7) (1:1000; cat. no. 9145, Cell Signalling); anti-STAT3 (D3Z2G) (1:1000, cat. no. 12640, Cell Signalling); anti-caspase 3 (D3R6Y) (1:1000, cat. no. 142200, Cell Signalling); anti-cleaved-caspase 3 (D175) (1:1000, cat. no. 9661, Cell Signalling); anti-cleaved-PARP (Asp 214) (1:1000, cat. no. 9541, Cell Signalling); AR TIC LE IN PR ES S anti-tubulin (DM1A) (1:200, cat. no. sc-32293, Santa Cruz Biotechnology) and anti-β-actin (AC-15) (1:5000, cat. no. 1978, Sigma-Aldrich).
Article Title: Nuclear actin and DNA replication stress regulate telomere maintenance by telomerase
Article Snippet: Antibodies used were anti-Chk1 (1:1000, Cell Signaling Technology, 2360S), anti-Chk1 (p345) (1:1000, Cell Signaling Technology, 2348S), anti-Vinculin (1:10000, Sigma-Aldrich, V9131), anti-Cofilin1 (1:1000, Cell Signaling Technology, 5175S), anti-Importin9 (1:1000, Thermo Fisher Scientific, PA141395 ), anti-Actin (1:2500, Sigma-Aldrich, A5441), anti-p70 (1:1000, Cell Signaling Technology, 2708S), anti-p70 (T389) (1:1000, Cell Signaling Technology, 9234S), anti-Histone 2 A (1:1000, Cell Signaling Technology, 12349S), anti-ATR (1:1000, Cell Signaling Technology, 2790S) and anti-ATRIP (1:1000, Thermo Fisher Scientific, PA1-519).
Western Blot:Article Title: Targeting de novo pyrimidine synthesis confers vulnerability to copper-mediated ATR inactivation in PARP inhibitor-resistant ovarian cancer.
Article Snippet: .. The following antibodies were used for western blot analysis: anti-p-ATR (S428) (1:1000; 2853, Cell Signaling Technology, MA, USA), anti-ATR (1:1000; 13934, Cell Signaling Technology), anti-p-CHK1 (S345) (1:1000; 2348, Cell Signaling Technology), anti-CHK1 (1:1000; ab40866, Abcam, Cambridge, UK), anti-ATRIP (1:1000; 2737, Cell Signaling Technology), anti-p-ATM (S1981) (1:1000; 13050, Cell Signaling Technology), anti-ATM (1:1000; 2873, Cell Signaling Technology), anti-p-CHK2 (T68) (1:1000; 2197, Cell Signaling Technology), anti-CHK2 (1:1000; 6334, Cell Signaling Technology), anti-γ-H2AX (1:5000; AP0687, ABclonal, Wuhan, China), anti-β-actin (1:4000; A5316, Sigma-Aldrich). .. The following antibodies were used for IF analysis: anti-p-ATR (S428) (1:100; AP0676, ABclonal), anti-ATR (1:200; ab313435, Abcam), anti-pCHK1 (S345) (1:50; 2348, Cell Signaling Technology), anti-CHK1 (1:200; ab40866, Abcam), anti-γ-H2AX (1:50; AP0687, ABclonal).
Article Title: Targeting de novo pyrimidine synthesis confers vulnerability to copper-mediated ATR inactivation in PARP inhibitor-resistant ovarian cancer
Article Snippet: .. The following antibodies were used for western blot analysis: anti-p-ATR (S428) (1:1000; 2853, Cell Signaling Technology, MA, USA), anti-ATR (1:1000; 13934, Cell Signaling Technology), anti-p-CHK1 (S345) (1:1000; 2348, Cell Signaling Technology), anti-CHK1 (1:1000; ab40866, Abcam, Cambridge, UK), anti-ATRIP (1:1000; 2737, Cell Signaling Technology), anti-p-ATM (S1981) (1:1000; 13050, Cell Signaling Technology), anti-ATM (1:1000; 2873, Cell Signaling Technology), anti-p-CHK2 (T68) (1:1000; 2197, Cell Signaling Technology), anti-CHK2 (1:1000; 6334, Cell Signaling Technology), anti-γ-H2AX (1:5000; AP0687, ABclonal, Wuhan, China), anti-β-actin (1:4000; A5316, Sigma-Aldrich). .. The following antibodies were used for IF analysis: anti-p-ATR (S428) (1:100; AP0676, ABclonal), anti-ATR (1:200; ab313435, Abcam), anti-p-CHK1 (S345) (1:50; 2348, Cell Signaling Technology), anti-CHK1 (1:200; ab40866, Abcam), anti-γ-H2AX (1:50; AP0687, ABclonal).
Colony Assay:Article Title: AURKA inhibitor VIC-1911 induces mitotic defects and functional BRCAness, sensitizing prostate cancer to PARP inhibition.
Article Snippet: After washing, the blots were incubated with HRP-conjugated secondary antibodies for 1h and visualized using the ECL substrate (Amersham) and a ChemiDoc imaging system (Bio-Rad). .. The following antibodies were used: anti-pAurora A(Thr288)/B(Thr232)/C(Thr198) (CST, 2914), anti-pTACC3 (Ser558) (CST, 8842), anti-TACC3 (CST, 8069), anti-Aurora A (CST, 91590), anti-pATM (Ser1981) (Abclonal, AP1030), anti-pKAP1(Ser824) (CST, 4127), anti-Actin (Santa Cruz, SC-47778), anti-Cleaved caspase 3 (CST, 9664S), anti-phospho-γH2AX (S139) (Abcam, ab2893), anti-cleaved PARP (CST, 5625S), anti-GAPDH (CST, 2118S), anti-p21 (CST, 2947S), anti- ERK1/2 (Santa Cruz, sc-398015), anti-pERK1/2 (Thr202/Tyr204) (CST, 9101), antipAKT (CST, 4060), anti-AKT (CST, 4691), anti-RAD51 (Calbiochem, PC130), anti-pATR (T1989) (GeneTex, GTX128145), anti-ATR (CST, 2851), anti-Vinculin (Abcam, ab129002), Cell proliferation (IncuCyte), cell viability, colony-formation assays Cell proliferation was monitored using the IncuCyte Live-Cell Imaging System (Sartorius Biotech), following the manufacturer’s instructions. ..
Live Cell Imaging:Article Title: AURKA inhibitor VIC-1911 induces mitotic defects and functional BRCAness, sensitizing prostate cancer to PARP inhibition.
Article Snippet: After washing, the blots were incubated with HRP-conjugated secondary antibodies for 1h and visualized using the ECL substrate (Amersham) and a ChemiDoc imaging system (Bio-Rad). .. The following antibodies were used: anti-pAurora A(Thr288)/B(Thr232)/C(Thr198) (CST, 2914), anti-pTACC3 (Ser558) (CST, 8842), anti-TACC3 (CST, 8069), anti-Aurora A (CST, 91590), anti-pATM (Ser1981) (Abclonal, AP1030), anti-pKAP1(Ser824) (CST, 4127), anti-Actin (Santa Cruz, SC-47778), anti-Cleaved caspase 3 (CST, 9664S), anti-phospho-γH2AX (S139) (Abcam, ab2893), anti-cleaved PARP (CST, 5625S), anti-GAPDH (CST, 2118S), anti-p21 (CST, 2947S), anti- ERK1/2 (Santa Cruz, sc-398015), anti-pERK1/2 (Thr202/Tyr204) (CST, 9101), antipAKT (CST, 4060), anti-AKT (CST, 4691), anti-RAD51 (Calbiochem, PC130), anti-pATR (T1989) (GeneTex, GTX128145), anti-ATR (CST, 2851), anti-Vinculin (Abcam, ab129002), Cell proliferation (IncuCyte), cell viability, colony-formation assays Cell proliferation was monitored using the IncuCyte Live-Cell Imaging System (Sartorius Biotech), following the manufacturer’s instructions. ..
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