Cycloheximide hy 12320

Manufactured by MedChemExpress

Sourced in United States

Cycloheximide (HY-12320) is a chemical compound used in various laboratory applications. It functions as a protein synthesis inhibitor, specifically targeting the eukaryotic 60S ribosomal subunit. This action interrupts the translocation step in protein synthesis, thereby preventing the formation of functional proteins.

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Lab products found in correlation

Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Dmem f12 basic medium, by Thermo Fisher Scientific (1 mentions) Bafilomycin a1, by Selleck Chemicals (1 mentions) Chloroquine diphosphate, by Merck Group (1 mentions) Gsk lsd1 2hcl, by Selleck Chemicals (1 mentions) Sp2509, by Selleck Chemicals (1 mentions) 6 well culture dishes, by NEST Biotechnology (1 mentions) Z vad fmk, by Selleck Chemicals (1 mentions) Mg132, by Selleck Chemicals (1 mentions) Rapamycin, by Merck Group (1 mentions) Dbcamp, by Merck Group (1 mentions) 8 br cgmp, by Merck Group (1 mentions) Mdl 12 330a, by Merck Group (1 mentions) L name, by Beyotime (1 mentions) L nmma, by Beyotime (1 mentions) Mg132, by Beyotime (1 mentions) Lipo6000 transfection reagent, by Beyotime (1 mentions) Anti normal rabbit igg, by Cell Signaling Technology (1 mentions) Protein a g magnetic beads hy k0202, by MedChemExpress (1 mentions) Du145, by American Type Culture Collection (1 mentions)

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HY-12320 (36 citations)

8 protocols using cycloheximide hy 12320

Ovarian Tissue Culture with Inhibitors

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Ovaries were separated by microdissection from the mesonephros or ovarian capsule in prechilled PBS (10 mM, pH 7.4) under a stereomicroscope (ZSA302, COIC). The isolated ovaries were cultured in 6‐well culture dishes (NEST Biotechnology) with basic DMEM/F‐12 medium (Gibco, Life Technologies) and Penicillin‐Streptomycin at 37°C in a 5% CO₂, 95% air atmosphere with saturated humidity.
Ovaries were cultured in either medium with dimethylsulfoxide (DMSO) or medium supplemented with inhibitor. The concentration of inhibitors used in this study: GSK‐LSD1‐2Hcl (S7574; Selleck): 64 μM; 3MA (S2767; Selleck, USA): 2.5 mM; Z‐VAD‐fmk (S7023; Selleck): 50 μM; Chloroquine diphosphate (C6688; Sigma): 10 μM; Bafilomycin A1 (S1413; Selleck): 60 nM; MG132 (S2619; Selleck): 5 μM; Cycloheximide (HY‐12320; MedChemExpress): 2 nM; and SP2509 (S7680; Selleck): 5 nM.

He M., Zhang T., Zhu Z., Qin S., Wang H., Zhao L., Zhang X., Hu J., Wen J., Cai H., Xin Q., Guo Q., Lin L., Zhou B., Zhang H., Xia G, & Wang C. (2020). LSD1 contributes to programmed oocyte death by regulating the transcription of autophagy adaptor SQSTM1/p62. Aging Cell, 19(3), e13102.

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Pharmacological Modulation of cGMP/mTOR Signaling

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The NO donor SNAP (‘SN’, N3398), the sGC agonist YC-1 (‘YC’, Y102), the sGCs inhibitor ODQ (O3636), the cGMP analog 8-Br-cGMP (‘8Br’, B1381), the mTOR inhibitor rapamycin (R8781), the cAMP donor dbcAMP (D0260) and the adenylyl cyclase inhibitor MDL-12,330A (‘MDL’, M182) were all purchased from Sigma-Aldrich (USA). The PKG inhibitor KT5823 (‘KT’, S1691), the NOS inhibitor L-NMMA (‘NM’, S0011), the eNOS inhibitor L-NAME (‘NA’, S0006), the iNOS inhibitor SMT (‘SM’, S0008), the nNOS inhibitor spermidine (‘SP’, S0010) and the proteasome inhibitor MG-132 (‘MG132’, S1748) were purchased from Beyotime (China). The KIT inhibitor Axitnib (‘AX’, S1005) was purchased from Selleck (China). The protein synthesis inhibitor cycloheximide (HY-12320) was purchased from MedChemExpress (USA).

Zhao P., Song Z., Wang Y., Cai H., Du X., Li C., Lv J., Liu X., Guo M, & Chen Z. (2020). The endothelial nitric oxide synthase/cyclic guanosine monophosphate/protein kinase G pathway activates primordial follicles. Aging (Albany NY), 13(1), 1096-1119.

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Antibody Validation for Protein Studies

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Anti-UHRF1 (A2343), Anti-β-Tubulin (A12289), Anti-NCAM1 (A7913), Anti-Synaptophysin (A6344), Anti-USP7 (A13564), Anti-BTRC (A18232) were purchased from ABclonal (Wuhan,HuBei, China). Anti-Cleaved-PARP, Anti-AKT (phospho S473) were purchased from Abcam (Cambirdge,MA,USA). Anti- Akt (pan) (#4685), Anti-HA Tag (#3724), Anti-p21(#2947), Anti-Phospho-AKT (Thr308) (#13038), Anti-Normal Rabbit IgG (#2729) were purchased from Cell Signaling Technology(Danvers, MA, USA), Anti-AKT1 antibody [HL1145] (#GTX636416) was purchased from GenTex (Irvine, CA, USA), Anti-His Tag(A00186) was purchased from GenScript (Nanjing, Jiangsu, China), p-Thr(H-2) was purchased from Santa Cruze Biotechnology(Shanghai, China). Monoclonal ANTI-Flag® M2 antibody (F1804) and Normal Mouse IgG (12-371) were purchased from Merck (Darmstadt, Germany).
Cycloheximide (HY-12320), MK2206 (HY-108232), and Protein A/G Magnetic Beads (HY-K0202) were purchased from MedChemExpress LLC. (Shanghai, China). MG132 (T2154) was purchased from Topscience Co. Ltd. (Shanghai, China). Lipo6000™ Transfection Reagent(C0526), a cationic liposome for transfection of HEK293T cells, was purchased from Beyotime (Shanghai, China).

Fu Y., Cao T., Zou X., Ye Y., Liu Y., Peng Y., Deng T., Yin L, & Li X. (2023). AKT1 regulates UHRF1 protein stability and promotes the resistance to abiraterone in prostate cancer. Oncogenesis, 12(1), 1.

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Prostate Cancer Cell Culture Protocol

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RPMI 1640 medium, fetal bovine serum (FBS), and penicillin-streptomycin were purchased from Gibco (Grand Island, NY). The protein synthesis inhibitor cycloheximide (HY-12320), proteasome inhibitor MG132 (HY-13259), DNA transcription inhibitor actinomycin D (HY-17559), and Bobcat339 (HY-111558A) were obtained from Medchemexpress (Monmouth Junction, NJ). The detailed information about antibodies used in this study was shown in Table S1. PC3, DU145, and LNCaP cells were obtained from ATCC, and were cultured in RPMI 1640 medium supplemented with 10% FBS and 1% penicillin-streptomycin.

Chen Y., Pan C., Wang X., Xu D., Ma Y., Hu J., Chen P., Xiang Z., Rao Q, & Han X. (2021). Silencing of METTL3 effectively hinders invasion and metastasis of prostate cancer cells. Theranostics, 11(16), 7640-7657.

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Antibody-Based Protein Detection Assay

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The antibodies used in this study included anti-nsP4 IgG (PA5-117443, rabbit; Invitrogen, Carlsbad, USA), anti-β-actin IgG (4970S, rabbit; Cell Signaling Technology, Beverly, USA), anti-phospho (Ser51) eIF2α IgG (3398S, rabbit; Cell Signaling Technology), anti-eIF2α IgG (5324S, rabbit; Cell Signaling Technology), anti-Hsp90 IgG (ab203126, rabbit; Abcam, Cambridge, USA), anti-GST IgG (11213-MM01, mouse; Sino Biological, Beijing, China), anti-RACK1 IgG (5432S, rabbit; Cell Signaling Technology), and anti-ubiquitin IgG (ab134953, rabbit; Abcam). The secondary antibodies included HRP-conjugated AffiniPure goat anti-rabbit IgG (H+L) (SA00001-2; Proteintech, Wuhan, China) and HRP-conjugated AffiniPure goat anti-mouse IgG (H+L) (SA00001-1; Proteintech). MG132 (HY-13259), Harringtonolide (HY-N10335), and cycloheximide (HY-12320) were purchased from MedChemExpress (MCE, Monmouth Junction, USA).

Yan Y., Zhang F., Zou M., Chen H., Xu J., Lu S, & Liu H. (2024). Identification of RACK1 as a novel regulator of non-structural protein 4 of chikungunya virus. Acta biochimica et biophysica Sinica.

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Cell Line Culture and Compound Treatments

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Human NSCLC cell lines A549, H1299, H1975, H1650, HCC827, PC9, and H226 and human leukemia K562 cells were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and cultured in RPMI 1640 medium (Invitrogen, USA). All cells were cultured at 37 °C and 5% CO₂. All cell lines have been validated by STR profiling within the last three years and were tested for mycoplasma and microbial contamination.
Recombinant human EGF was purchased from PeproTech (PeproTech, USA). MG132 (S2619), Gefitinib (S1025), and AZD9291 (S7297) were from Selleck (Selleck, USA). Cycloheximide (HY-12320) and Chloroquine (HY-17589A) were from MedChemExpress (MCE, USA).

Shen J., Liu G., Qi H., Xiang X, & Shao J. (2023). JMJD5 inhibits lung cancer progression by facilitating EGFR proteasomal degradation. Cell Death & Disease, 14(10), 657.

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Investigating Oncolytic HSV-1 Efficacy in OSCC Cell Lines

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OSCC lines SCC9 and SCC25 were obtained from the BeNa Culture Collection (BNCC, China), SCC7 was gifted from Prof. Bernard Roizman at the University of Chicago. All these cells were cultured in DMEM (Thermo Scientific, USA) supplemented with 10% fetal bovine serum (FBS; Thermo Scientific, USA). Vero (CCL-81) cells were purchased from the American Type Culture Collection (USA) and cultured in DMEM supplemented with 50% newborn calf serum (NBCS; Thermo Scientific, USA). All cell lines were incubated at 37°C in a humidified atmosphere of 5% CO₂. Oncolytic HSV-1 recombinant virus T1012G (oHSV-1 T1012G) was previously described.¹⁴ Immunoglobulin (Ig)G (S20043008) was obtained from the Shenzhen Weiguang Biological Co., Ltd, China. Cycloheximide (HY-12320) and GSK2643943A (HY-111458) were purchased from MedChem Express (MCE, USA).

Lu R., Wu G., Chen M., Ji D., Liu Y., Zhou G.G, & Fu W. (2021). USP18 and USP20 restrict oHSV-1 replication in resistant human oral squamous carcinoma cell line SCC9 and affect the viability of SCC9 cells. Molecular Therapy Oncolytics, 23, 477-487.

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Characterization of Autophagy Regulation

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Osimertinib (TAGRISSO) was obtained from Astra Zeneca. Spautin-1 (S7888), 3-Methyladenine (S2767) and rapamycin (S1039) were purchased from Selleck. Cycloheximide (C6628) was purchased from Sigma-Aldrich, and Cycloheximide (HY-12320) from MedChem Express. Anti-LC3II (#12741S), SQSTM1/p62 (#8025S), Atg5 (#9980S), Beclin 1 (#3495S), phospho-(Ser93)-beclin1 (#14717S), Sox2 (#3579S), ALDH1A1 (#36671S), GAPDH (#2118S) antibodies were from Cell Signaling Technology.

Li L., Wang Y., Jiao L., Lin C., Lu C., Zhang K., Hu C., Ye J., Zhang D., Wu H., Feng M, & He Y. (2019). Protective autophagy decreases osimertinib cytotoxicity through regulation of stem cell-like properties in lung cancer. Cancer letters, 452.

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