Human rhabdomyosarcoma (RD; CCL-136; ATCC) cells were maintained in Dulbecco’s modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS; Life Technologies) at 37°C in a 5% CO2 incubator. Echovirus 11 strain (NCBI Accession No. OP764694) was isolated from a feces sample of a 24-day-old female neonate with enterovirus infection after passaging in the RD cells. RD cells were infected with echovirus at various multiplicity of infection (MOI) for 1.5 h in serum-free DMEM. The cells were washed with phosphate-buffered saline (PBS) and cultured in a completely fresh medium for various times as indicated until they were harvested. For autophagy induction experiments, cells were infected or mock-infected with echovirus for 1.5 h, then cultured in a complete medium containing rapamycin (Selleck, AY-22989) at indicated concentrations for the indicated times. For autophagy inhibition experiments, cells were cultured in DMEM containing indicated concentrations of 3-methyladenine (3-MA) (Selleck, S2767) for 2 h, followed by echovirus infection for 1.5 h, and then incubated with fresh DMEM for 16 h. Cell counting kit-8 (CCK8) (Vazyme, A311-01) assay was performed to examine the cytotoxicity of rapamycin or 3-MA to RD cells.
Ay 22989
Manufactured by Selleck Chemicals
Sourced in United States
AY-22989 is a laboratory equipment item. It is used for general laboratory applications. The core function of AY-22989 is to perform specific laboratory tasks.
Automatically generated - may contain errors
Lab products found in correlation
Ccl 136, by American Type Culture Collection (1 mentions)
S2767, by Selleck Chemicals (1 mentions)
A311 01, by Vazyme (1 mentions)
Ap24534, by Selleck Chemicals (1 mentions)
Jnk in 8, by Selleck Chemicals (1 mentions)
Pkf118 310, by Merck Group (1 mentions)
Afuresertib gsk2110183, by Selleck Chemicals (1 mentions)
Cu cpt9a, by InvivoGen (1 mentions)
Wortmannin ky12420, by Selleck Chemicals (1 mentions)
Pd0325901, by Selleck Chemicals (1 mentions)
Torin1, by Merck Group (1 mentions)
4 protocols using ay 22989
1
Echovirus 11 Infection in RD Cells
2
Memory CD4+ T Cell Modulation
3
Comprehensive Pharmacological Toolkit for Cell Biology
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PD0325901 (mirdametinib), PRT062607 (P505-15) HCl, R788 (fostamatinib), wortmannin (KY 12420), afuresertib (GSK2110183), AY-22989 (Rapamycin), and JNK-IN-8 were purchased from Selleck Chemical. PKF118-310 was purchased from Sigma-Aldrich. Torin 1 and ST2825 were purchased from Calbiochem/Merck Millipore and ChemScence, respectively.
Stable isotope-labeled nucleosides, G (13C10, 98%; 15N5, 96–98%), U (13C9, 98%; 15N2, 96–98%), A (13C10, 98%; 15N5, 96–98%), C (13C9, 98%; 15N3, 96–98%), and dG (13C10, 98%; 15N5, 96–98%) were purchased from Cambridge Isotope Laboratories for the quantification of nucleosides by LC-MS/MS. The EdU used in the in vivo proliferation assay was purchased from Tokyo Chemical Industry Co.
DSR-139970 (Cpd7), a TLR7 inhibitor, was kindly provided by Sumitomo Pharma Co., Ltd. CU-CPT9a, a specific TLR8 inhibitor, and R848 were purchased from InvivoGen. SRBC used in the phagocytic assay were obtained from Cosmo Bio Co.
RNA9.2s (20mer, UsGsUsCsCsUsUsCsAsAsUsGsUsCsCsUsUsCsAsA), ssRNA40 (GsCsCsCsGsUsCsUsGsUsUsGsUsGsUsGsAsCsUsC), PolyU (19mer, UsUsUsUsUsUsUsUsUsUsUsUsUsUsUsUsUsUsU), and ODN1668 (20mer, dTsdCsdCsdAsdTsdGsdAsdCsdGsdTsdTsdCsdCsTdsdGsdAsdTsdGsdCsdT), in which “s” depicts a phosphothioate linkage, were synthesized by FASMAC.
Stable isotope-labeled nucleosides, G (13C10, 98%; 15N5, 96–98%), U (13C9, 98%; 15N2, 96–98%), A (13C10, 98%; 15N5, 96–98%), C (13C9, 98%; 15N3, 96–98%), and dG (13C10, 98%; 15N5, 96–98%) were purchased from Cambridge Isotope Laboratories for the quantification of nucleosides by LC-MS/MS. The EdU used in the in vivo proliferation assay was purchased from Tokyo Chemical Industry Co.
DSR-139970 (Cpd7), a TLR7 inhibitor, was kindly provided by Sumitomo Pharma Co., Ltd. CU-CPT9a, a specific TLR8 inhibitor, and R848 were purchased from InvivoGen. SRBC used in the phagocytic assay were obtained from Cosmo Bio Co.
RNA9.2s (20mer, UsGsUsCsCsUsUsCsAsAsUsGsUsCsCsUsUsCsAsA), ssRNA40 (GsCsCsCsGsUsCsUsGsUsUsGsUsGsUsGsAsCsUsC), PolyU (19mer, UsUsUsUsUsUsUsUsUsUsUsUsUsUsUsUsUsUsU), and ODN1668 (20mer, dTsdCsdCsdAsdTsdGsdAsdCsdGsdTsdTsdCsdCsTdsdGsdAsdTsdGsdCsdT), in which “s” depicts a phosphothioate linkage, were synthesized by FASMAC.
4
Investigating PI3K-VMP1 Relationship
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3-Methyladenine (3-MA, S1039)—a PI3K inhibitor—and rapamycin (AY-22989, S2767)—a PI3K activator—were used to investigate the relationship between PI3K and VMP1. Myoblast cells were incubated with 3-MA or AY-22989 for 3 h as described by Zhang et al. [36 (link)]. A CCK-8 assay was used to determine the optimal concentrations of 3-MA and AY-22989 for use in the experiments. The 3-MA and AY-22989 were purchased from Selleck Chemicals (Houston, TX, USA).
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