kinase 6 cdk6  (Proteintech)

Journal: Cureus

Article Title: Sterile Alpha Motif Domain-Containing 5 Suppresses Malignant Phenotypes and Tumor Growth in Breast Cancer: Regulation of Polo-Like Kinase 1 and c-Myc Signaling in a Xenograft Model

doi: 10.7759/cureus.73259

Figure Lengend Snippet: (A)-(B) MDA-MB-231 and HCC1937 cells were co-transfected with Sterile Alpha Motif Domain-Containing 5 (SAMD5) overexpression (OE) and Polo-like Kinase 1 (PLK1) OE and examined for the protein levels of MYC Proto-Oncogene (c-Myc), β-catenin, Cyclin-Dependent Kinase 4 (CDK4), Cyclin-Dependent Kinase 6 (CDK6), and Cyclin D1 using immunoblotting. **p < 0.01 vs. vector group; ##p < 0.01 vs. SAMD5 group

Article Snippet: The primary antibodies used were as follows: anti-SAMD5 (STJ194894; St John’s Laboratory, London, UK), anti-Ki67 (28074-1-AP; Proteintech), anti-Matrix Metallopeptidase 2 (MMP2) (10373-2-AP; Proteintech), anti-Matrix Metallopeptidase 9 (MMP9) (82854-1-RR; Proteintech), anti-PLK1 (ab189139; Abcam), anti-β-Catenin (51067-2-AP; Proteintech), anti-Cyclin-Dependent Kinase 4 (CDK4) (11026-1-AP; Proteintech), anti-Cyclin-Dependent Kinase 6 (CDK6) (14052-1-AP; Proteintech), anti-Cyclin D1 (26939-1-AP; Proteintech), anti-Cyclin E (11935-1-AP; Proteintech), and anti-GAPDH (60004-1-Ig; Proteintech).

Techniques: Transfection, Sterility, Over Expression, Western Blot, Plasmid Preparation

Journal: Cureus

Article Title: Sterile Alpha Motif Domain-Containing 5 Suppresses Malignant Phenotypes and Tumor Growth in Breast Cancer: Regulation of Polo-Like Kinase 1 and c-Myc Signaling in a Xenograft Model

doi: 10.7759/cureus.73259

Figure Lengend Snippet: (A) A xenograft tumor model was established in nude mice, and lentivirus-mediated overexpression of Sterile Alpha Motif Domain-Containing 5 (SAMD5) and/or Polo-like Kinase 1 (PLK1) was introduced into the tumors. (B)-(C) Tumor weight and volume were measured. (D) The protein levels of MYC Proto-Oncogene (c-Myc), β-catenin, Cyclin-Dependent Kinase 4 (CDK4), Cyclin-Dependent Kinase 6 (CDK6), and Cyclin D1 in tumor tissues were determined using immunoblotting. **p < 0.01 vs. vector group; #p < 0.05; ##p < 0.01 vs. SAMD5 group

Article Snippet: The primary antibodies used were as follows: anti-SAMD5 (STJ194894; St John’s Laboratory, London, UK), anti-Ki67 (28074-1-AP; Proteintech), anti-Matrix Metallopeptidase 2 (MMP2) (10373-2-AP; Proteintech), anti-Matrix Metallopeptidase 9 (MMP9) (82854-1-RR; Proteintech), anti-PLK1 (ab189139; Abcam), anti-β-Catenin (51067-2-AP; Proteintech), anti-Cyclin-Dependent Kinase 4 (CDK4) (11026-1-AP; Proteintech), anti-Cyclin-Dependent Kinase 6 (CDK6) (14052-1-AP; Proteintech), anti-Cyclin D1 (26939-1-AP; Proteintech), anti-Cyclin E (11935-1-AP; Proteintech), and anti-GAPDH (60004-1-Ig; Proteintech).

Techniques: Over Expression, Sterility, Western Blot, Plasmid Preparation

Journal: Cell Communication and Signaling : CCS

Article Title: Characterization of two melanoma cell lines resistant to BRAF/MEK inhibitors (vemurafenib and cobimetinib)

doi: 10.1186/s12964-024-01788-3

Figure Lengend Snippet: Changes in proliferation rate and cell cycle phases distribution in resistant melanoma cell lines. A Melanoma WM9 and Hs294T cells were seeded on a 96-well plate, and their proliferation rate was calculated as a ratio of the spectrophotometric signal after 48 h divided by the signal at t0. B Cell cycle analysis in WM9 and Hs294T melanoma control cells and cells resistant to treatment with BRAFi/MEKi. Western blot analysis of the level of proteins involved in cell cycle regulation: ( C ) CDK6, ( D ) p18, ( E ) p21, and ( F ) p27 in cell lysates obtained from control and resistant melanoma cells. The signal was normalized to the total protein content assessed by Ponceau S staining. Representative blotting membranes of three independent experiments are shown. Control (CTRL) constitutes WM9 and Hs294T cells treated with regular media with DMSO at the concentration used for drug delivery. Cells for cell cycle analysis as well as those used for Western blotting analysis were synchronized to obtain reliable results for cells dividing at widely varying rates as described in the ‘Materials and Methods’ section. The graphs show average values from at least three independent experiments ± SD. Asterisks in the graphs indicate statistical significance (p) at the level of * ≤ 0.05, ** ≤ 0.01, *** ≤ 0.001, and **** ≤ 0.0001

Article Snippet: Primary antibodies were directed against pERK1/2 (phosphorylated extracellular signal-regulated kinases 1/2) (Cell Signaling Technologies, #9101, 1:1000), AKT (Protein kinase B) (Cell Signaling Technologies, #4691, 1:1000), pAKT (phosphorylated protein kinase B) (Cell Signaling Technologies, #9271, 1:1000), p38 (p38 mitogen-activated protein kinase) (Cell Signaling Technologies, #8690, 1:1000), p-p38 (phosphorylated p38 mitogen-activated protein kinase) (Cell Signaling Technologies, #4511, 1:1000), JNK (c-Jun N-terminal kinase) (Santa Cruz Biotechnology, sc-7345, 1:200), p-JNK (phosphorylated c-Jun N-terminal kinase) (Santa Cruz Biotechnology, sc-6254, 1:200), CYP1A1 (Cytochrome P450 family 1 subfamily A member 1) (Santa Cruz Biotechnology, sc-25304, 1:200), ALCAM (CD166 antigen) (Santa Cruz Biotechnology, sc-74558, 1:200), TGFβRIII (Transforming growth factor-beta receptor III) (Cell Signaling Technologies, #2519, 1:1000), TGFβRI (Transforming growth factor-beta receptor I) (Santa Cruz Biotechnology, sc-398, 1:200), SOX2 (SRY-box transcription factor 2) (Cell Signaling Technologies, #3578, 1:1000), CDK6 (cyclin-dependent kinase 6) (Cell Signaling Technologies, #3136, 1:1000), p18 (CDKN2C (cyclin dependent kinase inhibitor 2 C) (Cell Signaling Technologies, #2896, 1:1000), p21 ((CDKN1A) cyclin dependent kinase inhibitor 1 A) (Cell Signaling Technologies, #2947, 1:1000), and p27 ((CDKN1B) cyclin dependent kinase inhibitor 1B) (Cell Signaling Technologies, #3686, 1:1000).

Techniques: Cell Cycle Assay, Control, Western Blot, Staining, Concentration Assay