Normal rat kidney fibroblast cells (NRK-49F, from American Type Culture Center, Manassas, VA) were cultured in Dulbecco’s Modified Eagle Medium supplemented with 2 mM L-glutamine, 1% non-essential amino acids and 5% fetal bovine serum (Gibco, Carlsbad, CA) in a 5% CO2, 37 °C humidified incubator. To examine the pro- or anti-fibrotic effects of TGF-β1 and TSA, subconfluent cultures of NRK-49F cells were first maintained quiescent in basal medium with 0.5% fetal bovine serum for 24 hrs, and then stimulated with 5 ng/ml TGF-β1 (PeproTech, Rocky Hill, NJ) in the presence or absence of TSA (Sigma-Aldrich, St Louis, MO). To investigate the roles of MAPKs and Notch signaling pathways in TGF-β1-mediated fibrogenesis, subconfluent NRK-49F cells were treated with the following inhibitors: 10 μM SB203580 ( p38 inhibitor; Gibco, Frederick, MD), 20 μM PD98059 (ERK inhibitor; Gibco, Frederick, MD), 10 μM SP600125 (JNK inhibitor; Gibco, Camarillo, CA) and RO4929097 (γ-secretase inhibitor; Selleckchem, Houston, TX).
Nrk 49f
Manufactured by Thermo Fisher Scientific
Sourced in United States
The NRK-49F is a laboratory instrument manufactured by Thermo Fisher Scientific. It is a multipurpose device designed for various applications in research and analysis. The NRK-49F is capable of performing a range of tasks, but a detailed description of its core function and intended use is not available at this time.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
Tgf β1, by Thermo Fisher Scientific (6 mentions)
Nrk 49f, by American Type Culture Collection (3 mentions)
Ro4929097, by Selleck Chemicals (2 mentions)
Sp600125, by Thermo Fisher Scientific (2 mentions)
Pd98059, by Thermo Fisher Scientific (2 mentions)
Sb203580, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Basic fgf, by Thermo Fisher Scientific (1 mentions)
P1399, by Merck Group (1 mentions)
C11995500bt, by Thermo Fisher Scientific (1 mentions)
Insulin, by Merck Group (1 mentions)
Nrk 52e, by R&D Systems (1 mentions)
Nrk 49f, by R&D Systems (1 mentions)
Recombinant human tgf β1, by R&D Systems (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Nrk 49f, by National Collection of Authenticated Cell Cultures (1 mentions)
Nrk 52e, by National Collection of Authenticated Cell Cultures (1 mentions)
Mycoblue mycoplasma detector, by Vazyme (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
8 protocols using nrk 49f
1
Investigating Fibrotic Effects of TGF-β1 and TSA
2
Isolation and Immortalization of TNC-Expressing Cells
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The TNC-expressing cells (TNC-Cell) were obtained by sorting the tdTomato-positive cells in the UUO kidneys of TNCCreER2-eGFP/+;R26tdTomato/+ mice using flow cytometry, and immortalized using SV40 T lentivirus (Fig. 9B ). This cell line was cultured in polylysine (20 μg/mL, Sigma, P1399) coated dish with DMEM/F12 media containing 10% FBS, 1 ng/mL basic-FGF (Peprotech, 400-29) and 5 ng/mL insulin (Sigma, I0305000). The normal rat kidney fibroblast NRK49F was purchased from American Type Culture Collection (ATCC). Both NRK49F and mouse embryo fibroblast NIH3T3 were cultured in DMEM media (GIBCO, C11995500BT) containing 10% heat-inactivated fetal bovine serum (FBS, GIBCO, 10270-106), 100U/ml penicillin and 100 μg/ml streptomycin. Mycoplasma contamination was tested by polymerase chain reaction (PCR). Cells were incubated at 37 °C in a 5% CO2, humidified atmosphere.
3
NRK49F Cells Fibrosis Regulation
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The renal fibroblast cell line NRK49F was purchased from Cell Culture Centre, Chinese Academy of Medical Sciences and Peking Union Medical College (Beijing, China). NRK49F cells were cultured in DMEM-Ham’s medium (Gibco, NY, United States) supplemented with 10% FBS (Gibco, NY, United States). The cells, after reaching approximately 80% confluence, were pre-treated with LM49 for 4 h, followed by incubation with recombinant TGF-β1 (5 ng/ml) for the indicated time.
4
Evaluating ABA's Effects on Kidney Cell Lines
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The normal rat kidney proximal tubular epithelial cell line (NRK-52E) and normal rat kidney interstitial fibroblast line (NRK-49F) purchased from the China Center for Type Culture Collection were used to evaluate the therapeutic effects of ABA on TGF-β1 or β-catenin stimulation. NRK-49F and NRK-52E cells were cultured in DMEM-F12 with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA) at 37°C with 5% CO2. Recombinant human TGF-β1 (R&D Systems, Minneapolis, MN, USA) was used at 5 ng/mL and 2.5 ng/mL, respectively, to stimulate NRK-49F and NRK-52E cells, in the presence or absence of ABA (10 μM). Recombinant human angiotensin II protein (R&D systems) was used at 1.0 µM and 2.0 µM to respectively stimulate NRK-49F and NRK-52E cells in the presence or absence of ABA (10 μM).
5
Cellular Transitions and Microcystin-RR Intervention
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The cell lines, HK-2 (a human proximal tubular cell line derived from the normal kidneys), and NRK-49F (a rat fibroblast cell line derived from the normal kidneys) were purchased from the Cell Bank of the Typical Culture Preservation Committee, Chinese Academy of Sciences. Both cells were tested negative for mycoplasma contamination using a MycoBlueTMmycoplasma detector (Vazyme, Nanjing, China). The cells were cultured in humidified air at 37°C with 5% CO2. HK-2 was grown in a DMEM/F12 medium (11330032, Gibco, New York, United States) and NRK-49F in DMEM medium (11965092, Gibco). All mediums were supplemented with 10% fetal bovine serum (10100147C, Gibco), 100 U/ml penicillin, and 100 μg/ml streptomycin. To induce epithelial to mesenchymal transition and fibroblast to myofibroblast transition in vitro, HK-2 and NRK-49F cell lines were grown in 6-well plates at a seeding density of 1×105 cells/ml in the medium with TGF-β1 (5 ng/ml) for 48 h. MC-RR (0.1 μM) was synchronously added to observe the intervention effect on the induced cells.
6
Cultivation and Treatment of Human and Rat Cell Lines
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Human proximal tubule cell line (HK-2) and rat fibroblast cell line (NRK-49F) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). HK-2 cells were grown in keratinocyte serum-free medium (Gibco, Waltham, MA, USA), supplemented with 50 µg/mL bovine pituitary extract and 5 ng/mL human recombinant epidermal growth factor. NRK-49F cells were cultured in Dulbecco’s modified Eagle medium/nutrient mixture F-12 medium (Gibco) with 5% fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin (Welgene, Gyeongsan, Korea). Cells were cultured in an atmosphere of 5% CO2 at 37°C in a humidity chamber and treated with 5 and 2 ng/mL of TGF-β1 (R&D Systems, Minneapolis, MN, USA), respectively, for the indicated time periods.
7
Cell Culture Protocols for Fibroblast Models
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Normal rat kidney fibroblasts (NRK-49F, ATCC, Manassas, VA, USA) and mouse embryonic fibroblasts (NIH-3T3, ATCC) were grown in Dulbecco’s modified Eagle’s medium (DMEM, Thermo Fisher Scientific, Waltham, MA, USA) containing 5% fetal bovine serum (FBS) and 1% antibiotics [29 (link),30 (link)]. Human embryonic kidney (HEK) 293T (ATCC) cells were cultured in DMEM with 10% FBS and 1% antibiotics. The cells were maintained in an incubator at 37 °C with 5% CO2. The NRK-49F and NIH-3T3 cells were incubated overnight in starvation media containing DMEM and 0.1% FBS before TGF-β1 (Peprotech, Rocky Hill, NJ, USA) treatment.
8
TGF-β1-Induced Renal Fibroblast Activation
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Normal rat renal fibroblasts NRK-49F cells, obtained from American Type Culture Collection (ATCC, Maryland, USA), were cultured in Dulbecco’s modified Eagle’s medium (Gibco, USA) containing 10% fetal bovine serum (Hyclone, USA), 100 μg/mL streptomycin and 100 U/mL penicillin. To test the effect of AN-FTS on TGF-β1-induced activation of NRK-49F, the cells were exposed to free FTS or AN-FTS (20 uM of FTS) containing 2 ng/mL of TGF-β1 (Peprotech, USA) for 48 h, then harvested for analysis.
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