Quercetin (Batch No. Z100081) was purchased from the National Institute for Food and Drug Control (Beijing, China). The leaves of Cacumen Platycladi were purchased from Nanjing Tongrentang (Nanjing, China). Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), and trypsin were purchased from BasalMedia (Shanghai, China). LPS and ATP were purchased from Sigma-Aldrich.
Fetal bovine serum (fbs)
Manufactured by BasalMedia
Sourced in China
FBS is a cell culture supplement derived from the blood of fetal bovine. It provides a complex mixture of proteins, growth factors, and other nutrients that support the growth and proliferation of various cell types in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by BasalMedia (4 mentions)
Dulbecco modified eagle medium (dmem), by BasalMedia (3 mentions)
Dulbecco s modified eagle medium dmem, by BasalMedia (2 mentions)
Dulbecco s modified eagle s medium, by BasalMedia (2 mentions)
Streptomycin, by BasalMedia (2 mentions)
Penicillin, by BasalMedia (2 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Trypsin, by BasalMedia (1 mentions)
Bovine serum albumin (bsa), by Maclin Biochemical Technology (1 mentions)
Protein loading buffer, by Ipsen (1 mentions)
Antibodies against ki67, by Cell Signaling Technology (1 mentions)
Hoechst 33342, by Yuanye Bio-Technology (1 mentions)
Antibodies against bax, by Cell Signaling Technology (1 mentions)
Calcein pi live dead assay kit, by Beyotime (1 mentions)
Sds page, by Ipsen (1 mentions)
Penicillin streptomycin, by Solarbio (1 mentions)
Dab kit, by ZSGB-BIO (1 mentions)
Trypsin edta, by Solarbio (1 mentions)
Dmem medium, by BasalMedia (1 mentions)
Jetprime, by Polyplus Transfection (1 mentions)
15 protocols using fetal bovine serum (fbs)
1
Quercetin Extraction and Cell Culture
2
Synthesis and Evaluation of Near-Infrared Dye
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Indocyanine green (ICG), human serum albumin (HSA, ≥98%), bovine serum albumin (BSA, ≥98%) were purchased from Macklin Inc. (Shanghai, China), Dulbecco’s modified Eagle’s medium and fetal bovine serum (FBS) were purchased from BasalMedia (Shanghai, China), trypsin-EDTA, penicillin-streptomycin were supplied by Solarbio (Shanghai, China). 3-(4,5-diMethylthialzol-2-yl)-2,5-diphe-nyltetrazolium bromide (MTT) and Hoechst33342 were purchased from YuanYe Bio-Technology (Shanghai, China). The universal SP kit (mouse/rabbit streptavidin–biotin detection system), DAB kit and PBS buffer (pH 7.4) were purchased from Zsbio (Beijing, China). All-purpose Powerful Antigen Retrieval Solution and Calcein/PI Live/Dead Assay kit and antibodies against Bcl-2 were purchased from Beyotime Institute of Biotechnology (Shanghai, China). Protein Loading Buffer, Multicolor Prestained Protein Ladder and 10% SDS-PAGE were from EpiZyme (Shanghai, China). Near-infrared dye IR-817 was synthesized following our previously reported protocol.26 (link) Antibodies against Ki-67 was purchased from Cell Signaling Technology (CST, Beverly, MA, USA). Antibodies against Cleaved caspase-3 (CC-3) was purchased from Zen-bio (Chengdu, China). Antibodies against Bax was purchased from Cell Signaling Technology (CST, Beverly, MA, USA).
3
Cell Culture and Transfection Protocol
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GES‐1 and MGC‐803 cells were cultured in RPMI‐1640 medium (BasalMedia) supplemented with 10% FBS (BasalMedia). AGS cells were cultured in Ham's F‐12 medium (BasalMedia) supplemented with 10% FBS. HEK293T and HGC27 cells were cultured in DMEM medium (BasalMedia) supplemented with 10% FBS. All cells were kept at 37°C in a 5% CO2 humidified incubator. JetPRIME® (Polyplus) was used to transfect cells according to the manufacturer's instructions.
4
Silencing FMNL1 in Renal Cell Carcinoma
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In this study, four human cell lines 786-O, Caki-1, HK2 and SNU349 were obtained from Shanghai cell Data. HK2 were cultured in DMEM (L110KJ, L210809, Basal Media), and 786-0, Caki-1, SNU349 were cultured in PRMI 1640 (L210KJ, B210904, Basal Media) with 10% fetal bovine serum (FBS; ABW) and 1% penicillin-streptomycin in a humidified atmosphere containing 5% CO2 at 37 °C. The shRNA sequences targeting FMNL1 and negative control sequence were designed and purchased from Shanghai Genechem Co., Ltd. 293T cells were used to generate lentivirus by co-transfecting GV493-shFMNL1s and negative control (CON313, hU6-MCS-CBh-gcGFP-IRES-puromycin) with helper plasmids pHelper 1.0 and pHelper 2.0. The lentiviral knockdown plasmid GV493-shFMNL1s were constructed and used to infect the 786-O and Caki-1 cell lines. The shRNA sequences targeting FMNL1 were listed as follows: shRNA1: ATCCAGACTAAGTTCCGAA, shRNA2: GCGGTTTCAAGTCAAGAAT, negative control sequence: TTCTCCGAACGTGTCACGT. Stable cell lines were selected by puromycin (786-O cells 2.5ug/ml, Caki-1 2ug/ml).
5
Investigating CHCHD4 Regulation in LUAD Cells
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Four human LUAD cell lines, NCI-H1975, H1299, A549, and H23, and the human bronchial epithelioid cell line 16-HBE, were supplied by Procell (Wuhan, China). All cell lines were maintained in RPMI-1640 medium (Hyclone) supplemented with 10% FBS (S660JJ, BasalMedia, Shanghai, China) and 1% streptomycin/penicillin (P1400, Solarbio, Beijing, China) in a humidified atmosphere of 5% CO2 at 37 °C. Small interfering RNAs against CHCHD4 (si-CHCHD4-1, si-CHCHD4-2, and si-CHCHD4-3) and their negative control (si-NC) were purchased from RiboBio (Guangzhou, China). Short hairpin RNAs (shRNAs) for CHCHD4 (sh-CHCHD4), and shRNA negative control (sh-NC) were obtained from Genechem (Shanghai, China). The pcDNA3.1-USF1 vector, pcDNA3.1-MYC vector and their empty vector were constructed by Tsingke Biotechnology Co., Ltd. Transfection was performed using Lipofectamine 3000 (Invitrogen, USA) according to the manufacturer's instructions.
6
AGE-Induced Keratinocyte Model for DFU
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Advanced glycation end products (AGE)-induced keratinocytes were applied to construct the in vitro DFU model. HaCaT cells (Human keratinocyte cells) were purchased from Shanghai Sunteam Biotechnology Co., Ltd (CAT: sunH8751; Shanghai, China) and routinely cultured in Dulbecco’s Modified Eagle’s medium (DMEM, BasalMedia, Shanghai, China) added with 10% fetal bovine serum (FBS, BasalMedia) and 1% Penicillin-Streptomycin (BasalMedia) at 37 °C cell incubator with 5% CO2. When the cells grew to approximately 30–40% confluence, they were treated with 60 mg/mL AGE-HSA, and then, cells in log phase were taken for further analysis.
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Advanced glycation end products (AGE)-induced keratinocytes were applied to construct the in vitro DFU model. HaCaT cells (Human keratinocyte cells) were purchased from Shanghai Sunteam Biotechnology Co., Ltd (CAT: sunH8751; Shanghai, China) and routinely cultured in Dulbecco’s Modified Eagle’s medium (DMEM, BasalMedia, Shanghai, China) added with 10% fetal bovine serum (FBS, BasalMedia) and 1% Penicillin-Streptomycin (BasalMedia) at 37 °C cell incubator with 5% CO2. When the cells grew to approximately 30–40% confluence, they were treated with 60 mg/mL AGE-HSA, and then, cells in log phase were taken for further analysis.
7
Cell Culture Protocols for Human and Mouse Cell Lines
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Detailed information for reagents and materials, including antibodies and cell lines, used in this study is provided in Supplementary Table 1 . Human DLD-1 cells were grown in McCoy’s 5A medium (BasalMedia) supplemented with 10% fetal bovine serum (FBS, Yeasen), 1× penicillin–streptomycin (Gibco). HEK293T cells and mouse embryonic fibroblasts (MEFs) were cultured with Dulbecco’s modified Eagle medium (DMEM, BasalMedia) supplemented with 10% FBS and 1× penicillin–streptomycin. HEK Expi293 cells were grown in suspension in serum-free medium. All cells were cultured at 37 °C and 5% CO2 and were negative for mycoplasma contamination.
8
Andrographolide-Induced Apoptosis in Lung Cancer
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Human lung adenocarcinoma cells (A549, H1299), human lung squamous cells (SK-MES-1), murine Lewis lung carcinoma cells (LLC), and normal human bronchial epithelial cells (BEAS-2B) were obtained from the American Type Culture Collection (Manassas, VA, United States). These cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, BasalMedia, Shanghai, China) which contains 10% fetal bovine serum (FBS, BasalMedia, Shanghai, China) and 1% penicillin–streptomycin solution (BasalMedia, Shanghai, China) at 37°C with 5% CO2. Andrographolide (Sigma-Aldrich, Germany) was dissolved into a concentration of 100 mM with dimethyl sulfoxide (DMSO, Sigma-Aldrich, Germany) and stored at -20°C. Antibody against β-actin was obtained from HuaBio (Hangzhou, China); Antibodies against cleaved-PARP (c-PARP), PARP, cleaved-caspase-3 (c-Casp3), caspase3 (Casp3), ATF4, c-Myc, Noxa, Puma, Bid, Bim, Bik, Bax, and Bak were all obtained from Cell Signaling Technology (Beverly, MA, United States).
9
Isolation and Senescence of Mouse Aortic VSMCs
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As described before,19 (link) primary VSMCs from the mouse aorta were extracted. To obtain primary VSMCs, thoracoabdominal aortas were extracted from healthy C57BL6 mice. The collected tissues were washed with DMEM (BasalMedia, Shanghai, China) and cut into 1 mm3 pieces. The pieces were plated onto 24-well plates and digested with 1 mg/mL collagenase (Sigma-Aldrich) and 0.5 mg/mL elastase (Sigma-Aldrich) for 1.5 h at 37°C. Collected VSMCs were cultured in DMEM with 100 U/mL penicillin, streptomycin, and 10% FBS (BasalMedia) in a humidified atmosphere containing 5% CO2. Cells from passages to 3–5 were used for subsequent analyses. VSMCs were treated with 50 μg/mL oxidized low-density lipoprotein (ox-LDL; Solarbio, China) for 72 h of incubation at 37°C to induce VSMC senescence.
10
Gastric Cell Lines Cultured in DMEM
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GC cell lines (SGC-790, SNU216, BGC823, and HGC27) and a gastric epithelial cell line (GES-1) were purchased from the American Type Culture Collection (Manassas, VA, USA) and were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (Basal Media, Shanghai, China) and penicillin-streptomycin (BasalMedia, Shanghai, China). All of the cells were incubated in a humidified air atmosphere with 5% CO2 at 37°C. LY294002, a PI3-kinase inhibitor (ab120243), was used to treat cells for 2 hrs at 30 µM.
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