Fetal bovine serum (FBS), trypsin-EDTA (0.25%), Pierce™ BCA Protein Assay kit, FxCycle™ PI/RNase Staining Solution and Dulbecco's modified Eagle's medium (DMEM) were purchased from Thermo Fisher Scientific, Inc. L-15 medium containing 80 U/ml penicillin and 80 µg/ml streptomycin (Nanjing KeyGen Biotech Co., Ltd.) was used to culture MDA-MB-231 cells. Western and IP Lysis Buffer were acquired from Beyotime Institute of Biotechnology. CCK-8 and antibody for glyceraldehyde 3-phosphate dehydrogenase (GAPDH; polyclonal; cat. no. ABP57259) were obtained from Abbkine, Inc. Matrigel was purchased from BD Biosciences. Cyclin B1 polyclonal (product no. 4138S) antibody and β-actin monoclonal (product no. 4970S) antibody were purchased from Cell Signaling Technology, Inc. CDK1 monoclonal (product code ab18) antibody was purchased from Abcam. PNO1 antibodies were purchased from LSBio, Inc. (polyclonal; cat. no. LS-C179090) and Santa Cruz Biotechnology, Inc. (monoclonal; cat. no. sc-514727) respectively. Goat anti-rabbit IgG HRP-conjugated secondary antibody (cat. no. L3012) and goat anti-mouse HRP-conjugated IgG secondary antibody (cat. no. L3032) were purchased from Signalway Antibody LLC.
L15 medium
Manufactured by Keygen Biotech
Sourced in China
L15 medium is a cell culture medium designed to support the growth and maintenance of a variety of cell types. It is a balanced salt solution that provides the necessary nutrients, vitamins, and other components required for cell survival and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Mccoy s 5a medium, by Keygen Biotech (14 mentions)
Sw620, by Keygen Biotech (9 mentions)
Fetal bovine serum (fbs), by Sartorius (8 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (7 mentions)
Hct116, by Keygen Biotech (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Sw480, by Keygen Biotech (5 mentions)
Dld 1, by Thermo Fisher Scientific (4 mentions)
Ht 29, by Thermo Fisher Scientific (4 mentions)
Hct116, by Thermo Fisher Scientific (4 mentions)
Sw620, by National Collection of Authenticated Cell Cultures (4 mentions)
Dld 1, by National Collection of Authenticated Cell Cultures (4 mentions)
Hct116, by National Collection of Authenticated Cell Cultures (4 mentions)
Sw480, by National Collection of Authenticated Cell Cultures (4 mentions)
Hct116, by Wuhan Boster Biological Technology (4 mentions)
Hct116, by Apexbio (3 mentions)
Sw620, by Apexbio (3 mentions)
Sr11302, by Apexbio (3 mentions)
Sw480, by Wuhan Boster Biological Technology (3 mentions)
Sw620, by Wuhan Boster Biological Technology (3 mentions)
19 protocols using l15 medium
1
Comprehensive Methodology for Cell Culture and Protein Analysis
2
Investigating c-Jun Phosphorylation in CRC Cells
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Human CRC cell lines (HCT116, SW620, SW480, DLD-1, HT-29) were purchased from the Cell Bank of the Chinese Academy of Science (Shanghai, China). The human normal colorectal epithelial cell line FHC was obtained from the American Type Culture Collection (Manassas, VA, USA). Cell lines were cultured in the appropriate medium supplemented with 10% fetal bovine serum (FBS; Gibco, NY, USA) and 1% antibiotic/antimycotic solution and maintained in an incubator at 37°C with 5% CO 2 in a humidi ed atmosphere. DLD-1 and HT-29 cells were maintained in RPMI-1640 medium (Gibco), FHC cells were cultured in Dulbecco's modi ed Eagle medium (DMEM; Gibco), HCT116 cells were maintained in McCoy's 5A medium (KeyGEN, Nanjing, China) and SW620 cells were maintained in L-15 medium (KeyGEN). To test the effects of the N-terminal phosphorylation of c-Jun, HCT116 and SW620 cells were pretreated with SR11302 (10 μM for 1 h) (APExBIO, USA) before transient transfection [30] .
3
Cell Culture Conditions for CRC Lines
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Human CRC cells SW620, HCT116, SW480, HT29, and normal colonic epithelia cells (NCM460) were purchased from Boster Company (Boster, Wuhan, People’s Republic of China). SW620 and SW480 were grown in L15 medium (KeyGEN BioTECH, Nanjing, China) supplemented with 10% FBS (Biological Industries, Israel). HCT116, HT29, and NCM460 cells were grown in McCoy′s5A medium (KeyGEN BioTECH, Nanjing, China) supplemented with 10% FBS (Biological Industries, Israel). All cells were incubated at 37 °C with 5% CO2.
4
Culturing Breast Cell Lines
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The MDA-MB-231 breast cancer cell line and the MCF10A healthy breast cell line were purchased from the American Type Culture Collection. Cells were cultured in L-15 medium (Nanjing KeyGen Biotech Co., Ltd.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 U/ml streptomycin (Nanjing KeyGen Biotech Co., Ltd.) in humidified 5% CO2 at 37°C.
5
Culturing Colorectal Cancer Cell Lines
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Colorectal cancer cell lines were purchased from American Type Culture Collection (ATCC). SW620 and SW480 cells were cultured in L15 medium (KeyGEN BioTECH, Nanjing, China) and supplemented with 10% fetal bovine serum (FBS; Bio-Channel, Nanjing, China). LoVo cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Carlsbad, CA, USA) and supplemented with 10% FBS. DLD-1 cells were cultured in RPMI 1640 medium (Roswell Park Memorial Institute 1640; Gibco, Carlsbad, CA, USA) and supplemented with 10% FBS. All cells used in this study were maintained in a 5% CO2 cell culture incubator at 37 °C.
6
FAK Inhibitor's Impact on Cell Signaling
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The BRACs used in this study were purchased from the New Star Natural Plant Development Company (Jilin, China). The FAK inhibitor, 1,2,4,5-benzenetetramine tetrahydrochloride (Y15), was obtained from Sigma (St. Louis, MO, USA). Fetal bovine serum (FBS) and horse serum were purchased from Gibco-BRL (Grand Island, NY, USA). Dulbecco's modified Eagle's medium (DMEM)/high glucose and DMEM/F12 were purchased from HyClone (Beijing, China). L15 medium was obtained from Keygen Biotech (Nanjing, China). Primary antibodies against FAK (ab40794), phospho-FAK (ab38458, ab81298), Src (ab16885), phospho-Src (ab24789), and p130Cas (ab136514) were purchased from Abcam (Cambridge, MA, USA). Phospho-p130Cas (#4011) antibody was purchased from Cell Signaling Technology (Danvers, MA, USA). Primary antibodies against E-cadherin (610181), vimentin (550513), and fibronectin (610077) were purchased from BD Biosciences (Bedford, MA, USA).
7
Cell Culture Conditions for Human Colorectal Cancer
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Human CRC cell lines (SW480, SW620, HCT116, HT29) and normal colonic epithelial cell lines (NCM460) were purchased from Wuhan Boster Biological Technology, Ltd. (Wuhan, China). SW480 and SW620 cells were cultured in L15 medium (Nanjing KeyGen Biotech Co., Ltd., Nanjing, China) containing 10% fetal bovine serum (FBS: Biological Industries Israel Beit-Haemek, Beit-Haemek, Israel). HCT116 and HT29 were cultured with McCoy's 5A medium (Nanjing KeyGen Biotech Co., Ltd.) containing 10% FBS. NCM460 was cultured in Dulbecco's Modified Eagle Medium (DMEM, Thermo Fisher Scientific™, Beijing China) containing 10% FBS. All cells were incubated at 37 °C in a 5% CO2 incubator.
8
Culture and Characterization of Human CRC Cell Lines
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Human CRC cell lines (SW620 and SW480) were cultured in L15 medium (KeyGEN BioTECH, Nanjing, China) supplemented with 10% fetal bovine serum (FBS) (Biological Industries, Israel), and HCT116 and HT-29 cells were cultured in McCoy‘s 5A medium (KeyGEN BioTECH, Nanjing, China) supplemented with 10% FBS. Cells were grown in a 5% CO2 cell culture incubator at 37 °C. Clinical samples were obtained from patients treated at the Third Xiangya Hospital of Central South University (Hunan, China) under informed consent and approval by the Ethics Committee of Central South University (more details about the clinical samples can be found in Table 1 ).
9
Colorectal Cancer Cell Line Cultivation
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Human CRC cell lines (HCT116, SW620, SW480, DLD-1, HT-29) were purchased from the Cell Bank of the Chinese Academy of Science (Shanghai, China). The human normal colorectal epithelial cell line FHC was obtained from the American Type Culture Collection (Manassas, VA, USA). Cell lines were cultured in the appropriate medium supplemented with 10% fetal bovine serum (FBS; Gibco, NY, USA) and 1% antibiotic/antimycotic solution and maintained in an incubator at 37 °C with 5% CO2 in a humidified atmosphere. DLD-1 and HT-29 cells were maintained in RPMI-1640 medium (Gibco), FHC cells were cultured in Dulbecco’s modified Eagle medium (DMEM; Gibco), HCT116 cells were maintained in McCoy’s 5A medium (KeyGEN, Nanjing, China) and SW620 cells were maintained in L-15 medium (KeyGEN). To test the effects of the N-terminal phosphorylation of c-Jun, HCT116 and SW620 cells were pretreated with SR11302 (10 μM for 1 h) (APExBIO, USA) before transient transfection [30 ].
10
Colon Cancer Cell Lines and Culture
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Normal human colon mucosal epithelial (FHC) cells and HT29, HCT116, SW480, and SW620 colon cancer cells were purchased from Nanjing KeyGen Biotech Co., Ltd., China. The FHC cells were cultured in RPMI 1640 medium (Gibco, USA) supplemented with 10% foetal bovine serum (FBS, Biological Industries, Israel) and penicillin/streptomycin solution. The HT29 and HCT116 cells were cultured in McCoy’s 5A medium (Keygen Biotech) supplemented with 10% FBS. The SW480 and SW620 cells were cultured in L15 medium (KeyGen Biotech) containing 10% FBS. All the cells were cultured in a cell culture incubator at 37 °C with 5% CO2. All the cells were identified within the past year based on short tandem repeats (STRs) or single nucleotide polymorphisms (SNPs) and were not contaminated with mycoplasma.
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