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CAS:33419-42-0
Appearance:White to off-white solid
Storage:Powder : 2-8℃, 2 years; In solvent(mother liquid): -20℃, 1 month; -80℃, 6 months
Purity:HPLC≥98%
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Examples of using this product（for reference only）
In Vitro:
Cell（ U2OS, H209, H69, Hep3B, and H1299 cells，Etoposide: H69 3 μm, H209 5 μm, U2OS 10 μm, Hep3B 15 μm, H1299 15 μm）:
Cancer Cells with Chemotherapy Resistance Generation and Analysis: U2OS, H209, H69, Hep3B, and H1299 cells were treated with different concentrations of cisplatin (Solarbio) and etoposide (Solarbio, IE270). Subsequently, the cells were cultured for 1 month at a concentration with a lower apoptotic rate (Cisplatin: H69 2.5 μm, H209 4 μm, U2OS 7 μm, Hep3B 3 μm, H1299 3.5 μm; Etoposide: H69 3 μm, H209 5 μm, U2OS 10 μm, Hep3B 15 μm, H1299 15 μm). The drug-resistant cells were divided into two groups and treated with the same gradient concentration of the drug as before, one group was treated with 0.5 mm F1,6P at the same time. MTT assay was used to detect cell viability and calculate IC50.
Reference：
Li Y, Fu Y, Zhang Y, Duan B, Zhao Y, Shang M, Cheng Y, Zhang K, Yu Q, Wang T. Nuclear Fructose-1,6-Bisphosphate Inhibits Tumor Growth and Sensitizes Chemotherapy by Targeting HMGB1. Adv Sci (Weinh). 2023 Mar;10(7):e2203528. doi: 10.1002/advs.202203528. Epub 2023 Jan 15. PMID: 36642839; PMCID: PMC9982576.
Cell（HeLa cells and KGN cells，5 μg/mL ETO，1 h）:
HeLa cells and KGN cells were cultured in 24-well plates and transiently transfected with wild-type, mutant, and polymorphic HSF2BP-FLAG plasmids with the use of Lipofectamine 3000. After being cultured for 48 h, the cells were divided into three groups: without treatment, treated with etoposide(ETO; 5 μg/ml, Solarbio, Beijing, China) for 1 h, and treated with mitomycin (MMC; 500 ng/ml) for 8 h. Then, the treated cells were cultured with a normal medium and recovered for a specific time at 37℃.
Reference：
Li S, Xu W, Xu B, Gao S, Zhang Q, Qin Y, Guo T. Pathogenic Variations of Homologous Recombination Gene HSF2BP Identified in Sporadic Patients With Premature Ovarian Insufficiency. Front Cell Dev Biol. 2022 Jan 31;9:768123. doi: 10.3389/fcell.2021.768123. PMID: 35174157; PMCID: PMC8841426.
Cell Etoposide sensitivity assay（KGN cells，5 ng/μl etoposide，6 h）:
KGN cells were seeded in 6‐well plates and cultured 48 hr after transfection or infection. Cells were harvested immediately or exposed to 5 ng/μl etoposide (ETO; Solarbio, Beijing, China) for 6 hr to induce DNA damage, followed by harvesting immediately or after recovery for 1 hr and 2 hr. DNA damage marker γH2AX was detected in KGN cells with or without ETO treatment and recovery by western blot.Clonogenic survival（ HeLa cells，0.5 ng/μl ETO，6 h）HeLa cells were seeded in 6‐well plates and cultured 48 hr after transfection or infection. Then cells were exposed to 0.5 ng/μl ETO (Solarbio, Beijing, China) for 6 hr, followed by recovery for 2 hr. The cells were harvested and counted, then reseeded into a new 6‐well plate by 500 cells per well (three wells for duplicate) and incubated for 14 days. Colonies were stained with crystal violet and counted.
Reference：
Zhang X, Dang Y, Liu R, Zhao S, Ma J, Qin Y. MicroRNA-127-5p impairs function of granulosa cells via HMGB2 gene in premature ovarian insufficiency. J Cell Physiol. 2020 Nov;235(11):8826-8838. doi: 10.1002/jcp.29725. Epub 2020 May 11. PMID: 32391592.
In Vivo:
Mice（21‐day‐old female C57BL/6J mice，10 mg/kg ETO，I.P）：
The 21‐day‐old female C57BL/6J mice were purchased and maintained in a temperature‐ and humidity‐controlled room on a 1‐hr light/12‐hr dark cycle. The orthotopic ovarian mouse model was established as previously described. A total of 20 μl miR‐127‐5p mimics or controls at 1 nmol with Lipofectamine 2000 was injected into the left and right ovarian bursa by insulin syringes, respectively. Forty‐eight hours after injection, mice were treated with 10 mg/kg ETO (Solarbio, Beijing, China) intraperitoneally to induce DNA damage. After 6 hr, the ovaries were excised and ruptured for γH2AX detection.
Reference：
Zhang X, Dang Y, Liu R, Zhao S, Ma J, Qin Y. MicroRNA-127-5p impairs function of granulosa cells via HMGB2 gene in premature ovarian insufficiency. J Cell Physiol. 2020 Nov;235(11):8826-8838. doi: 10.1002/jcp.29725. Epub 2020 May 11. PMID: 32391592.
