Bafilomycin A1 (Autophagy inhibitor), XAV939 (β-catenin inhibitor) and SKL2001(β-catenin agonist) were from MedChemExpress.
Skl2001
Manufactured by MedChemExpress
Sourced in United States
SKL2001 is a laboratory centrifuge designed for high-speed separation of samples. It is capable of reaching speeds of up to 10,000 RPM, making it suitable for a variety of applications that require efficient separation of particles, cells, or other components from liquid samples.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Xav 939, by MedChemExpress (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Iwp 2, by Beyotime (2 mentions)
Bafilomycin a1, by MedChemExpress (1 mentions)
Fh535, by MedChemExpress (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Nci h1650, by American Type Culture Collection (1 mentions)
Tgf β1, by MedChemExpress (1 mentions)
Nrk 52e, by American Type Culture Collection (1 mentions)
Recombinant human tgf β1, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Arpe 19, by Solarbio (1 mentions)
Recombinant human tgf β1, by Novoprotein (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Arpe 19, by American Type Culture Collection (1 mentions)
Puromycin, by Solarbio (1 mentions)
Pcdna3.1 plasmid, by YouBio (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
12 protocols using skl2001
1
Autophagy and Wnt Signaling Modulation
2
Modulation of PBMSC Proliferation and Survival by β-catenin and Oct4
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To determine the effect of β-catenin on the proliferation and survival of PBMSCs, we randomly divided the cells into the following four groups according to different intervention methods: the no intervention group (serving as the control group), the β-catenin overexpression group (oeβ-catenin), the β-catenin knockdown group (shβ-catenin), and the β-catenin overexpression+β-catenin knockdown group (oeβ-catenin plus shβ-catenin). To determine the effect of Oct4 on the proliferation and survival of PBMSCs, we divided the cells into the no intervention group, the Oct4 overexpression group (oeOct4), the Oct4 knockdown group (shOct4), and the Oct4 overexpression+Oct4 knockdown group (oeOct4 plus shOct4). To evaluate the effects of β-catenin and Oct4 on cell growth and apoptosis, we added oeβ-catenin+shOct4 and shβ-catenin+oeOct4. After transfection, all groups of PBMSCs were cultured for 70 days for subsequent experiments to analyze the growth and apoptosis of PBMSCs.
To investigate the mechanisms of β-catenin-mediated cytoprotective effects against apoptotic cell death, we performed β-catenin activation in PBMSCs using oeβ-catenin or a β-catenin agonist (SKL2001, MedChemexpress, Monmouth Junction, NJ, USA), and β-catenin inhibition was carried out by shβ-catenin or a β-catenin inhibitor (FH535, MedChemexpress).
To investigate the mechanisms of β-catenin-mediated cytoprotective effects against apoptotic cell death, we performed β-catenin activation in PBMSCs using oeβ-catenin or a β-catenin agonist (SKL2001, MedChemexpress, Monmouth Junction, NJ, USA), and β-catenin inhibition was carried out by shβ-catenin or a β-catenin inhibitor (FH535, MedChemexpress).
3
Culturing and Treating NSCLC Cell Lines
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Human NSCLC cell lines (A549 and NCI-H1650) were obtained from the American Type Culture Collection (TCGA, Manassas, Virginia, USA). Cells were cultured in DMEM (Sigma‒Aldrich, Darmstadt, Germany) containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (Gibco, Grand Island, New York, USA) in a 5% CO2 incubator at 37 °C. The cells were cultured in medium containing drug-containing serum. For β-catenin agonist treatment, NSCLC cells were treated with 30 µM SKL2001 (MedChem Express, Monmouth Junction, NJ, USA) for 24 h.
4
Modulation of Wnt/β-catenin Pathway in NRK52E Cells
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NRK52E (ATCC, Manassas, VA, USA) cell line was cultured in a DMEM medium (Hyclone, Logan, UT). The cells were maintained at 37 °C under 5% CO2 in a humidified chamber. After incubation in the medium containing 1% FBS for 24 h, cells were treated with 10 ng/ml Recombinant Human TGF-β1 (Peprotech, NJ, USA) alone, 100 ng/ml relaxin alone or co-treated with TGF-β1 and relaxin for 48 h. To induce the Wnt/β-catenin pathway, NRK52E cells were treated with 40 μM SKL2001 (MedChemExpress LLC, Monmouth Junction, NJ, USA).
5
Modulating Wnt/β-catenin Signaling in DPCs
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SKL2001 (HY-101085, MedChemExpress, Shanghai, China) was used as an agonist of the Wnt/β-catenin signal pathway and IWP-2 (SF6831, Beyotime, Shanghai, China) was used as an inhibitor of the Wnt/β-catenin signal pathway [27 (link),28 (link)]. The DPCs were grown to 50–80% confluence and treated with 40 μΜ SKL2001 or 10 μΜ IWP-2 for 24 h.
6
Modulation of Hepatocellular Carcinoma Cell Lines
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Normal human hepatocyte (LO2) as well as HCC cells (HLE, Huh-7, HepG2 and SMMC-7721) were purchased from the Cell Culture Center, Chinese Academy of Medical Sciences (Shanghai, China). The cells were cultured using Dulbecco’s modified Eagle’s medium (DMEM; Gibco, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, USA) and incubated at 37°C in a humidified atmosphere with 5% CO2. Huh-7 and SMMC7721 cells were transfected with AQP9 overexpressing vector (LV-AQP9) or the control (LV-NC). Subsequently, the cells were selected with 1 μg/mL puromycin for four weeks. The cells were harvested 48 h post-transfection for use in further experiments. To inhibit or induce Wnt/β-catenin pathway, Huh-7 cells were treated with 10 mM XAV939 (MedChemExpress LLC, Monmouth Junction, NJ, USA), and SMMC-7721 were treated with 40 μM SKL2001 (MedChemExpress LLC, Monmouth Junction, NJ, USA), respectively.
7
Wnt/β-catenin Pathway Modulation in ARPE-19 Cells
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A human retinal pigment epithelial cell line (ARPE‐19) was purchased from American Type Culture Collection (ATCC, Manassas, VA, US) and cultured in Dulbecco's Modified Eagle's Medium/Nutrient Mixture F‐12 (DMEM/F‐12; Gibco, Grand Island, NY, US) supplemented with 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, US) at 37°C in a humid atmosphere containing 5% CO2. Recombinant human TGFβ1 was purchased from Novoprotein. SKL2001, the agonist of wnt/β‐catenin pathway, was purchased from MedChemExpress.
The pCDH‐Vec control vectors and pCDH‐METTL3 overexpression vectors as well as pLKO.1‐TRC‐Vec control vectors and pLKO.1‐TRC‐shMETTL3 knockdown vectors used for stable transfection were kindly provided by Prof. Shuibin Lin.18 Plasmids were transfected into ARPE‐19 cells using lentivirus, as described previously. Stably transformed ARPE‐19 cells were selected by selection on media containing 2 μg/ml puromycin (Solarbio Life Science, CN) for 48 h.28
The pCDH‐Vec control vectors and pCDH‐METTL3 overexpression vectors as well as pLKO.1‐TRC‐Vec control vectors and pLKO.1‐TRC‐shMETTL3 knockdown vectors used for stable transfection were kindly provided by Prof. Shuibin Lin.
8
Gastric Cancer Cell Culture and Transfection
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We selected several commonly used GC cell lines, SNU-1, HGC-27, MKN-7, MKN-45, MKN-74 and AGS, in recent years with different differentiations, and they were supplied by BeNa Culture Collection (Beijing, China), also was the normal human gastric mucosal epithelial cells (GES-1). We supplemented the RPMI-1640 medium (Sigma-Aldrich, Darmstadt, Germany) with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, Waltham, USA) and 1% penicillin/streptomycin (Gibco, Carlsbad, USA) for cell culture. The incubation environment was 5% CO2 and 37°C with saturated humidity.
Sequence design and synthesis services were provided by GeneChem (Shanghai, China). The pcDNA3.1 plasmid (YouBio, Changsha, China) was used to construct CXXC5 and KANK1 overexpression vectors. Based on Lipofectamine 3000 (Invitrogen, Waltham, USA), the transfection assay was performed with a concentration of 50 nM for small interfering RNA (siRNA) and 0.5 μg for overexpression plasmids. SKL2001 was purchased from MedChemExpress (New Jersey, USA) and was used to treat cells after transfection at a concentration of 40 μM.
Sequence design and synthesis services were provided by GeneChem (Shanghai, China). The pcDNA3.1 plasmid (YouBio, Changsha, China) was used to construct CXXC5 and KANK1 overexpression vectors. Based on Lipofectamine 3000 (Invitrogen, Waltham, USA), the transfection assay was performed with a concentration of 50 nM for small interfering RNA (siRNA) and 0.5 μg for overexpression plasmids. SKL2001 was purchased from MedChemExpress (New Jersey, USA) and was used to treat cells after transfection at a concentration of 40 μM.
9
Collagen I and Honokiol Modulate HTR-8/SVneo Cell Viability
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HTR-8SV/neo cells were seeded in six-well plate which were incubated with different concentration of collagen I for 48h. CCK-8 (ab228554, Abcam, MA, USA) assay was conducted to detect cell viability and cell cycle analysis cell cycle stage was analyzed using flow cytometry. Western blotting was used to measure the relative protein level. The experimental procedures were detailed in the Supplementary Material
Honokiol, which enhance the phosphorylation of ERK phosphorylation, (HY-N0003, MedChemExpress, USA) dissolved at a concentration of 10μM. In the co-treatment group, HTR-8/SVneo cells were treated with Honokiol and collagen I 48h. SKL-2001 (HY-101085, MedChemExpress, USA), an agonist of β-catenin which was used at a concentration of 5μM. Co-treament with collagen I 24h. Cell viability, cell cycle and relative protein level were analyzed by CCK-8, cycle cell and western blotting.
Honokiol, which enhance the phosphorylation of ERK phosphorylation, (HY-N0003, MedChemExpress, USA) dissolved at a concentration of 10μM. In the co-treatment group, HTR-8/SVneo cells were treated with Honokiol and collagen I 48h. SKL-2001 (HY-101085, MedChemExpress, USA), an agonist of β-catenin which was used at a concentration of 5μM. Co-treament with collagen I 24h. Cell viability, cell cycle and relative protein level were analyzed by CCK-8, cycle cell and western blotting.
10
Lung Cancer Cell Line Experimentation
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A549 and NCI-H460 human lung cancer cell lines were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA). The normal pulmonary epithelial cell line, BEAS-2B, was purchased from Cobioer (Nanjing, China). The cell lines were maintained in RPMI-1640 medium (Biological Industries, Kibbutz Beit Haemek, Israel) containing 10% fetal bovine serum (FBS) (Biological Industries, Kibbutz Beit Haemek, Israel) at 37°C with 5% CO2. The Wnt signaling agonist, SKL2001 (40 μM) was purchased from MedChem Express (Monmouth Junction, NJ, USA). Itraconazole was purchased from Selleck (Houston, TX, USA).
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