The mouse C2C12 myoblasts (ATCC, Manassas, USA) were cultured at 37°C with 5% CO2 in DMEM (ThermoFisher, Massachusetts, USA), 10% FBS (ThermoFisher, Massachusetts, USA), 2 mM L-glutamine, 100 IU/mL penicillin, and 100 μg/mL streptomycin (ThermoFisher Scientific). When the C2C12 myoblasts grew to 70–80% confluence, the culture medium was switched to a differentiation medium which consisted of DMEM, 2% horse serum (Thermofisher, Massachusetts, USA), 100 IU/mL penicillin, and 100 μg/mL streptomycin. After 5 days, most of the myoblasts differentiated into myotubes. Our previous studies had shown that the concentration of CSE below 3% had no significant effects on the viability of C2C12 myotubes,17 (link) so subsequent experiments took 3% as the optimal effective dose. In the cell culture, C2C12 myotubes were also treated with indicated agents, including transforming growth factor (TGF)-β receptor activing-like kinase (ALK) 4/5 blocker TEW-7197 (S7530, Selleck, Shanghai, China), ferroptosis inhibitor UAMC-3203 (S8792, Selleck, Shanghai, China), hypoxia-inducible factor (HIF) 2α (400087, Merk Millipore), recombinant MSTN (120–00-10, Dogesce, Beijing, China) and vehicle (dimethylsulfoxide, DMSO).
Uamc 3203
Manufactured by Selleck Chemicals
Sourced in China
UAMC-3203 is a laboratory instrument designed for the analysis and measurement of various chemical and biological samples. It utilizes advanced spectroscopic techniques to provide accurate and reliable data.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Tew 7197, by Selleck Chemicals (1 mentions)
C2c12 myoblasts, by Thermo Fisher Scientific (1 mentions)
Pd 146176, by Selleck Chemicals (1 mentions)
Z vad fmk, by Solarbio (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Liproxstatin 1, by Selleck Chemicals (1 mentions)
Necrostatin 1, by Merck Group (1 mentions)
Ferrostatin 1, by Merck Group (1 mentions)
Pgl3 basic vector, by Promega (1 mentions)
Z lehd fmk, by Selleck Chemicals (1 mentions)
Z ietd fmk, by Selleck Chemicals (1 mentions)
Mab0041, by R&D Systems (1 mentions)
Mab210, by R&D Systems (1 mentions)
Necrostatin 1, by Cayman Chemical (1 mentions)
Mab375, by R&D Systems (1 mentions)
4 protocols using uamc 3203
1
Differentiation of C2C12 Myoblasts
2
Cell Culture and Transfection Conditions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Ishikawa, HEK293T and H1299 cell lines were cultured in DMEM (Corning, 10-013-CVR) supplemented with 10% (v/v) FBS (Gibco, 10099141). The HEC-265 cell line was cultured in EMEM (M&C Gene Technology, CM10010) with 15% FBS. The AN3CA cell line was cultured in EMEM supplemented with 10% FBS, 1× NEAA (Gibco, 11140050) and 1 mM sodium pyruvate (M&C Gene Technology, CC007). The cell lines were originally purchased from ATCC or Cell Resource Center of IBMS-CAMS, freshly thawed from our stock and cultured for no longer than 2 months. All cell lines were negative for mycoplasma contamination. The transfections were conducted by using Lipofectamine 2000 (Invitrogen, 11668500) according to the manufacturer’s protocol. To generate a luciferase reporter, annealed oligos were cloned into a pGL3-basic vector (Promega, E1761). MPA was purchased from Selleck (S2567). The following reagents were used as: Z-VAD-FMK (Solarbio, IZ0050) 10 µg/ml; necrostatin-1 (Sigma‒Aldrich, N9037) 10 µg/ml; ferrostatin-1 (Sigma‒Aldrich, SML0583) 2 µM; 3-MA (Sigma‒Aldrich, M9281) 2 mM; PD146176 (Selleck, S6956) 5 µM; liproxstatin-1 (Selleck, S7699) 2 µM; UAMC-3203 (Selleck, S8792) 2 µM. The antibodies and the sequence of siRNA, sgRNA and primers are listed in Supplementary Table S1 .
3
Neuroprotective Effects of UAMC-3203 and DFO
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Animals were randomized into five groups (n = 6 per group): Sham, Control, UAMC-3203, DFO, UAMC-3203 + DFO. Sham underwent exactly the same surgical procedures as the other groups except without VF and CPR. Five groups were administered with UAMC-3203 (Selleckchem, S8792, 5 mg/kg, 1 mg/mL), DFO (Cayman, 14595, 100 mg/kg, 100 mg/mL) or/and corresponding placebo (2%DMSO, saline) by intraperitoneal injection at the start of PC. Investigators involved in CPR were blinded to the group assignments.
At 6 h after ROSC, the rats were euthanized by an intravenous injection of Euthasol (A commercial euthanasia solution containing pentobarbital sodium and phenytoin sodium, 150 mg/kg). The heart was rapidly harvested and frozen in liquid nitrogen for further assay. A routine necropsy was performed to determine whether there were gross abnormalities, including traumatic injuries caused by cannulation, airway management, surgical operation, or precordial compression.
At 6 h after ROSC, the rats were euthanized by an intravenous injection of Euthasol (A commercial euthanasia solution containing pentobarbital sodium and phenytoin sodium, 150 mg/kg). The heart was rapidly harvested and frozen in liquid nitrogen for further assay. A routine necropsy was performed to determine whether there were gross abnormalities, including traumatic injuries caused by cannulation, airway management, surgical operation, or precordial compression.
4
UVB Irradiation of Cells and Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse dorsal fur was removed via shaving and depilation with Veet (Reckitt Benckiser, UK). Mice were placed in restrainers with facial protection and treated once ±250mJ/cm2 UVB light using the UV-2 ultraviolet irradiation system (Tyler Research). UVB light was provided by cascade-phosphor ultraviolet generators that emit 310nm UVB radiation.
For irradiation of cultured cells at approximately 80% confluence, growth medium was replaced with PBS and cells were treated with 0-50mJ/cm2 UVB light as indicated in specific experiments. PBS was removed immediately following irradiation and replaced with growth medium. Cells were incubated at 37°C for the indicated times.
Inhibitors of cell death (Ac-YVAD-cmk, Sigma Aldrich SML0429; Necrostatin-1, Cayman Chemical 11658; GSK’872, Tocris Bioscience 64-921-0; Z-IETD-FMK, Selleckchem S7314; Z-LEHD-FMK, Selleckchem S7313; Z-VAD-FMK, Selleckchem S7023; UAMC-3203, Selleckchem S8972) were added 30 minutes prior to IFN treatment and again immediately following UVB exposure.
For neutralizing antibody experiments, N/TERTs were treated with isotype control (R&D MAB002, MAB0041) or neutralizing antibodies targeting TRAIL (100ng/ml; R&D MAB375), TNF-α (5µg/ml; R&D MAB210) or FasL (1µg/ml; R&D MAB126) immediately following UVB exposure.
For irradiation of cultured cells at approximately 80% confluence, growth medium was replaced with PBS and cells were treated with 0-50mJ/cm2 UVB light as indicated in specific experiments. PBS was removed immediately following irradiation and replaced with growth medium. Cells were incubated at 37°C for the indicated times.
Inhibitors of cell death (Ac-YVAD-cmk, Sigma Aldrich SML0429; Necrostatin-1, Cayman Chemical 11658; GSK’872, Tocris Bioscience 64-921-0; Z-IETD-FMK, Selleckchem S7314; Z-LEHD-FMK, Selleckchem S7313; Z-VAD-FMK, Selleckchem S7023; UAMC-3203, Selleckchem S8972) were added 30 minutes prior to IFN treatment and again immediately following UVB exposure.
For neutralizing antibody experiments, N/TERTs were treated with isotype control (R&D MAB002, MAB0041) or neutralizing antibodies targeting TRAIL (100ng/ml; R&D MAB375), TNF-α (5µg/ml; R&D MAB210) or FasL (1µg/ml; R&D MAB126) immediately following UVB exposure.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!