Journal: Cell Reports Medicine

Article Title: Co-targeting RANK pathway treats and prevents acquired resistance to CDK4/6 inhibitors in luminal breast cancer

doi: 10.1016/j.xcrm.2023.101120

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti Cyclin D1 (92G2) , Cell Signaling , Cat#2978; RRID:AB_2259616.

Techniques: Recombinant, shRNA, Blocking Assay, Alamar Blue Assay, Flow Cytometry, Isolation, Bicinchoninic Acid Protein Assay, TUNEL Assay, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Software

Journal: Biomolecules

Article Title: Low-Temperature Plasma-Activated Medium Inhibits the Migration of Non-Small Cell Lung Cancer Cells via the Wnt/ β -Catenin Pathway

doi: 10.3390/biom13071073

Figure Lengend Snippet: Effects of LTP treatment on the expression of Wnt/ β -catenin and VEGF-signaling-pathway-associated proteins in PC-9 cells. ( A ) WB bands of Wnt3A, β -Catenin, Cyclin D1, c-Jun, VEGFR2, and VEGF proteins. ( B ) Quantitative analysis of each protein expression level. Data represent the mean ± SD of three independent experiments. “ns” means no statistical difference. * p < 0.05, *** p < 0.001 with ANOVA compared with the control.

Article Snippet: Rabbit anti-N-Cadherin antibodies (AB Clone, Wuhan, China, 1:1000), rabbit anti-E-Cadherin antibodies (AB Clone, Wuhan, China, 1:1000), rabbit anti-Vimentin antibodies (AB Clone, Wuhan, China, 1:1000), rabbit anti-c-Jun antibodies (Cell Signaling, Danvers, MA, USA, 1: 1000), and rabbit anti-Cyclin D1 antibodies (Cell Signaling, Danvers, MA, USA, 1: 1000) were incubated overnight at 4 °C, respectively.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: GP ‐2250, a novel anticancer agent, inhibits the energy metabolism, activates AMP‐Kinase and impairs the NF‐kB pathway in pancreatic cancer cells

doi: 10.1111/jcmm.17825

Figure Lengend Snippet: Representative western blot (A) and quantitative analysis (B) of pAMPK and cyclin D1. pAMPK: Time dependency of AMPK phosphorylation at T172 following incubation of Panc Tul with GP‐2250 (250 μM).) Dose‐dependency of AMPK phosphorylation after incubation of BxPC3 cells+ for 24 h. Cyclin D1: Time‐dependent decrease of cyclin D1 following incubation of Panc Tul (500 μM GP‐2250) and BxPC3 cells (250 μM GP‐2250). ß‐Actin/HSP‐90 used as internal controls. NC, negative control.

Article Snippet: Thereafter, the membranes were blocked in EveryBlot Blocking Buffer (BIO RAD) according to the manufacturer's antibody specification protocol for 5 min and incubated overnight at 4°C with the respective primary antibody [P‐AMPKalpha Rabbit mAB (T172) #2535, P‐ACC (Ser79) Rabbit mAB #11818, P‐Raptor (Ser792) Rabbit mAB #2083, p53 Rabbit mAB #2527, mTOR Rabbit Ab #2972, Akt Rabbit Ab #9272, cyclin D1 Rabbit mAB #55506, beta‐actin Rabbit mAB #8457, α‐tubulin Rabbit mAb #2125, Bcl2 Rabbit mAb #4223 and BAX Rabbit AB #2772 (CST)] at 1:1000 dilution.

Techniques: Western Blot, Incubation, Negative Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: GP ‐2250, a novel anticancer agent, inhibits the energy metabolism, activates AMP‐Kinase and impairs the NF‐kB pathway in pancreatic cancer cells

doi: 10.1111/jcmm.17825

Figure Lengend Snippet: Proposed scheme of the alteration of cancer cell metabolism and NF‐κB inhibition by GP‐2250. Bold black arrows indicate metabolic pathways; red and green arrows indicate drug‐induced changes. By limiting the energy metabolism through the inhibition of hexokinase 2 and GAPDH, GP‐2250 induces an energy deficit in line with an impairment of the TCA cycle. The reduction of ATP triggers the activation of the energy‐deficit sensor AMPK. Its downstream events include the inhibition of mTOR, a major driver of tumour cell growth and potential impairment of fatty acid synthesis (FAS) through ACC‐1 inhibition. The inhibition of NF‐κB by GP‐2250 limits the rate of tumour cell proliferation through cyclin D1 downregulation. It also promotes apoptosis through downregulation of the anti‐apoptotic Bcl2. ROS contributes to the upregulation of the transcription factor p53, which supports apoptosis. AMPK, adenosine monophosphate‐dependent protein kinase; FAS, fatty acid synthesis; NF‐κB, nuclear factor kappa‐light‐chain‐enhancer of activated B cells; PDH, pyruvate dehydrogenase.

Article Snippet: Thereafter, the membranes were blocked in EveryBlot Blocking Buffer (BIO RAD) according to the manufacturer's antibody specification protocol for 5 min and incubated overnight at 4°C with the respective primary antibody [P‐AMPKalpha Rabbit mAB (T172) #2535, P‐ACC (Ser79) Rabbit mAB #11818, P‐Raptor (Ser792) Rabbit mAB #2083, p53 Rabbit mAB #2527, mTOR Rabbit Ab #2972, Akt Rabbit Ab #9272, cyclin D1 Rabbit mAB #55506, beta‐actin Rabbit mAB #8457, α‐tubulin Rabbit mAb #2125, Bcl2 Rabbit mAb #4223 and BAX Rabbit AB #2772 (CST)] at 1:1000 dilution.

Techniques: Inhibition, Activation Assay

Journal: The Prostate

Article Title: Inverse agonism at the Na/K-ATPase receptor reverses EMT in prostate cancer cells

doi: 10.1002/pros.24144

Figure Lengend Snippet: Antibodies and Reagents

Article Snippet: Antibody Property Supplier Catalogue number dilution α1 NKA antibody (α6F) Mouse monoclonal Developmental Studies Hybridoma Bank of University of Iowa (Iowa) a6f 1:1000 PhosphoSrc (Tyr419) antibody Rabbit polyclonal Invitrogen 44–660G 1:1000 c-Src B-12 antibody Mouse monoclonal Santacruz Biotechnology sc-8056 1:1000 Rat α1 NKA antibody Rabbit polyclonal Dr. T.A.Pressley (Texas Tech University, TX) Not applicable 1:1000 Anti-phosphotyrosine antibody, clone 4G10 Mouse monoclonal EMD Millipore 05–321 1:1000 c-Myc antibody Santacruz Biotechnology 1:1000 Anti-tubulin antibody Mouse monoclonal SIGMA T5168 1:2000 Cyclin D1 antibody Rabbit monoclonal Cell Signaling Technology 2978S 1:1000 Cyclin E1 antibody Rabbit monoclonal Cell Signaling Technology 20808S 1:1000 p53 antibody Mouse pantropic Calbiochem OP43 1:1000 p21 antibody Rabbit polyclonal Santacruz Biotechnology sc-397 1:1000 Phospho MAPK antibody Rabbit Cell Signaling Technology 9101 1:1000 ERK1/2 antibody Rabbit polyclonal Santacruz Biotechnology sc-94 1:1000 Phospho-FAK (Tyr576/7) antibody Rabbit polyclonal Cell Signaling Technology 3281S 1:1000 FAK antibody Rabbit polyclonal Cell Signaling Technology 3285S 1:1000 Phospho-FAK (Tyr 397) antibody Rabbit polyclonal Cell Signaling Technology 3283S 1:1000 Anti-Na+/K+-ATPase β1 antibody clone 464.8 Mouse monoclonal EMD Millipore 05–382 1:1000 E-cadherin (24E10) antibody Rabbit monoclonal Cell Signaling Technology 3195S 1:1000 Anti β-catenin antibody Mouse monoclonal BD Bioscience 610153 1:1000 ZO-1 antibody Rabbit polyclonal Thermo-Fisher Scientific 61–7300 1:1000 ZO-2 antibody Rabbit polyclonal Thermo-Fisher Scientific 38–9100 1:1000 Occludin antibody (OC-3F10) Mouse monoclonal Thermo-Fisher Scientific 33–1500 1:1000 SNAIL (C15D3) antibody Rabbit monoclonal Cell Signaling Technology 3879S 1:1000 TCF8/ZEB1 antibody Rabbit monoclonal Cell Signaling Technology 3396S 1:1000 Vimentin (D21H3) antibody Rabbit monoclonal Cell Signaling Technology 5741S 1:1000 MMP-2 antibody Rabbit monoclonal Cell Signaling Technology 87809S 1:1000 MMP-9 antibody Rabbit monoclonal Cell Signaling Technology 13667S 1:1000 PCNA antibody Mouse monoclonal Santacruz Biotechnology sc-56 1:2000 Lamin B antibody Goat polyclonal Santacruz Biotechnology sc-6216 1:1000 β actin antibody Mouse monoclonal Santacruz Biotechnology sc-47778 1:1000 SLUG (C19G7) antibody Rabbit monoclonal Cell Signaling Technology 9585S 1:1000 N cadherin (D4R1H) antibody Rabbit monoclonal Cell Signaling Technology 13116S 1:1000 Open in a separate window Antibodies and Reagents Rat α1 NKA- specific antibody (NASE) was a kind gift from Dr. T. A. Pressley (Texas Tech University, TX) All reagents were obtained from Sigma-Aldrich except FAK inhibitor (Cat. No. 324877; Millipore).

Techniques: