VERO cells, HeLa cells, CHO cells, and human foreskin fibroblasts (HFF) were purchased from the American Type Culture Collection. All cells were maintained at 37°C in a 5% CO2 atmosphere and grown in either Alpha MEM (with Earle’s salts without ribonucleosides, deoxyribonucleosides, and glutamine) or DMEM (with 4.5 g/l glucose and sodium pyruvate without l -glutamine; Corning Cellgro). Unless otherwise stated, media was supplemented with 10% (vol/vol) FBS, 2 mM l- glutamine (Corning Cellgro), and 100 U/ml penicillin plus 100 mg/ml streptomycin (Quality Biological). HeLa cells were transiently transfected using 2 µg plasmid DNA and the Amaxa Nucleofector solution R according to the manufacturer’s instructions using program I-013 (Lonza) or with 0.25 µg plasmid DNA and JetPrime reagent according to the manufacturer’s instructions (Polyplus-transfection SA). To engineer a stable cell line expressing GFP-Rab11A, the GFP-Rab11A plasmid was linearized with ApaL1 and transfected into VERO cells using 2 µg of linearized plasmid, the Amaxa Nucleofector solution R, and program V-01, according to the manufacturer’s instructions. Stable clones were selected with 800 µg/ml G418 sulfate (Corning Cellgro) in Alpha MEM with 20 mM Hepes and cloned in serial dilutions in 96-well plates.
Hela cells
Manufactured by Lonza
HeLa cells are a widely used immortal cell line derived from cervical cancer cells. They are commonly used in various research applications, such as cell biology, cancer research, and drug development. HeLa cells have the ability to continuously divide and proliferate, making them a valuable tool for researchers.
Automatically generated - may contain errors
Lab products found in correlation
Alpha mem, by Corning (1 mentions)
Hela cell, by American Type Culture Collection (1 mentions)
Vero cell, by American Type Culture Collection (1 mentions)
Jetprime reagent, by Polyplus Transfection (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
L glutamine, by Corning (1 mentions)
G418 sulfate, by Corning (1 mentions)
Cho cells, by American Type Culture Collection (1 mentions)
Amaxa nucleofector solution r, by Lonza (1 mentions)
Penicillin, by Quality Biological (1 mentions)
Streptomycin, by Quality Biological (1 mentions)
Hek293, by Lonza (1 mentions)
Epidermal growth factor (egf), by Lonza (1 mentions)
Genetrans 2, by MoBiTec (1 mentions)
Hmvec dlyad, by Lonza (1 mentions)
Amphotericin b, by Lonza (1 mentions)
Gentamicin, by Lonza (1 mentions)
Neon transfection system, by Thermo Fisher Scientific (1 mentions)
S0615, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
4 protocols using hela cells
1
Cell Culture Protocols for Diverse Cell Lines
2
Functional Assays with Endothelial Cells
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Human umbilical vein endothelial cells (HUVECs), human dermal microvascular endothelial cells (HDMECs), human mammary epithelial cells (HMECs), human umbilical artery endothelial cells (HUAEC), human embryonic kidney 293 cells (HEK 293), human dermal lymphatic microvascular endothelial cells (HMVEC-dLyAd), and HeLa cells were obtained from Lonza. All functional assays were performed with HUVECS which were grown at 37 °C in endothelial basal medium (EBM) supplemented with hydrocortisone (1 μg/ml), bovine brain extract (12 μg/ml), gentamicin (50 μg/ml), amphotericin B (50 ng/ml), epidermal growth factor (10 ng/ml) (Lonza), and 10% fetal bovine serum (FBS, Perbio). Transfections of siRNA were performed using the GeneTrans II (MoBiTec) reagent according to the manufacturer’s protocols.
Except for scratch-wound assays, all functional assays were performed on fibronectin-plated HUVECs. Briefly, cells were harvested, left 30 min in suspension to recover from trypsinization, and seeded onto fibronectin-coated dishes for 30 min.
Except for scratch-wound assays, all functional assays were performed on fibronectin-plated HUVECs. Briefly, cells were harvested, left 30 min in suspension to recover from trypsinization, and seeded onto fibronectin-coated dishes for 30 min.
3
HeLa Cell Culture and Transfection
4
Cell Culture and Transfection Protocol
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WT and Mbnl1,2 DKO MEFs were gifts from Maurice Swanson. HeLa cells (ATCC), MCF7 cells and WT and Mbnl1,2 DKO MEFs were grown in high-glucose DMEM (Lonza) supplemented with 10% fetal bovine serum (FBS) (Sigma Aldrich) and 1 × penicillin–streptomycin (Sigma Aldrich). HSkM (Life Technologies) cells were grown in HAM F-10 medium (Sigma Aldrich) supplemented with 20% FBS, 1 × penicillin–streptomycin, 0.39 µg/mL dexamethasone (Sigma Aldrich) and 10 ng/mL epidermal growth (Sigma Aldrich). All cells were grown at 37 °C and an atmosphere containing 5% CO2. Prior to transfection, the cells were plated in 6-well or 12-well plates and transfected at 50–60% confluence with plasmids using Lipofectamine 3000 (Thermo Fisher) or with siRNA or antisense oligonucleotides using Lipofectamine RNAiMAX (Thermo Fisher) according to the manufacturer’s protocol. Genes were knocked down with siRNAs against MBNL1, MBNL2, DDX5, DDX17 or control siRNA was used (siCtrl) [9 (link), 37 (link)] (Sigma Aldrich) at 50 nM. SSOs with 2′-O-methoxyethyl-phosphorothioate (2′MOE-PS) modification, SSO-e7 and SSO-Ctrl, were used at 25 nM. Cotransfection with 200 ng of the ATP2A1 e22 minigenes and 250 ng of pEGFP-MBNL1-41 expression vector was preceded by siRNA treatment and 4 h of incubation period. The cells were harvested 48 h after transfection. The siRNA and SSO sequences are listed in Suppl. Table S1.
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