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CAS:69-09-0
Appearance:White to off-white solid
Storage:Powder : 2-8℃, 2 years
Purity:HPLC≥98%
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Examples of using this product（for reference only）
In Vitro:
Cell（HepG2, 20 μg/mL, 1 h, 37℃）:
For initial study of cellular internalization, NPs-EPI was incubated with tumor spheroids at an EPI concentration of 8 μg/mL for 4 h. The tumor spheroids were grown as our previous reported procedure. Briefly, HepG2 cells were seeded in 96-well plates at a density of 8×103 per well precoated with 50 μL of 1% low melting point agarose. Cells were cultured to obtain multicellular spheroids (400–500 μm in diameter) for subsequent studies. After incubation with NPs-EPI, tumor spheroids were rinsed three times with cold PBS and processed for image analysis by TEM.To investigate the internalization pathways of NPs-EPI, HepG2 cells were seeded into 6-well plates with 2×105 cells per well and incubated for 24 hours. The cells were then pre-treated with different endocytic inhibitors (chlorpromazine hydrochloride 20 μg/mL, amiloride hydrochloride 200 μg/mL) at 37℃ for 1 hour. Subsequently, NPs-EPI (EPI 8 μg/mL) were added and incubated at 37℃ for 2 hours. Finally, the cells were washed three times with cold PBS and harvested for flow cytometry to determine the fluorescence intensity of EPI (Ex=488, Em=588). Cells pre-treated with PBS at 37℃ served as the positive control group. Internalization assays were also performed in the absence of endocytic inhibitors at 4℃ when indicated.
Reference：
Chen E, Han S, Song B, Xu L, Yuan H, Liang M, Sun Y. Mechanism Investigation of Hyaluronidase-Combined Multistage Nanoparticles for Solid Tumor Penetration and Antitumor Effect. Int J Nanomedicine. 2020 Aug 24;15:6311-6324. doi: 10.2147/IJN.S257164. PMID: 32922003; PMCID: PMC7458542.
Cell （100 μM Chlorpromazine，30 min）:
To investigate the endocytosis mechanism involved in the uptake process, cells were pre-treated with endocytic inhibitors including chlorpromazine (100 μM), genistein (200 μM), and amiloride (200 μM) for 30 min. Ten the cells were treated with 1 μg of FITC-labeled DMSNs for 6 h. In a control group, cells were not treated with the inhibitors prior to nanoparticle treatment. Cellular uptakes of nanoparticles were quantitated by fow cytometer as described above.
Reference：
Mo C, Lu L, Liu D, Wei K. Development of erianin-loaded dendritic mesoporous silica nanospheres with pro-apoptotic effects and enhanced topical delivery. J Nanobiotechnology. 2020 Mar 30;18(1):55. doi: 10.1186/s12951-020-00608-3. PMID: 32228604; PMCID: PMC7104482.
Cell（A549 cells and MDCK cells；5 μg/ mL Chlorpromazine）:
To investigate the uptake mechanism of virus-laden AFPs, cells were pretreated with NA enzyme (1 U/ml) at 37℃ for 2 hours and immediately incubated with diverse inhibitors in complete cell culture medium at 37℃ for 1 hour, followed by infection with virus-laden AFPs (50 μg/ml) for 24 hours. Chlorpromazine (5 μg/ ml; Solarbio, China) was used to inhibit clathrin-mediated endocytosis;methyl-β-cyclodextrin (20 μg/ml) was used to inhibit lipid raft–mediated endocytosis; amiloride (50 μg/ml; Solarbio, China) was used to block micropinocytosis; cytochalasin D (2.5 μg/ml) was used to inhibit actin polymerization and macropinocytosis, as established previously.
Reference：
Dong Z, Ma J, Qiu J, Ren Q, Shan Q, Duan X, Li G, Zuo YY, Qi Y, Liu Y, Liu G, Lynch I, Fang M, Liu S. Airborne fine particles drive H1N1 viruses deep into the lower respiratory tract and distant organs. Sci Adv. 2023 Jun 9;9(23):eadf2165. doi: 10.1126/sciadv.adf2165. Epub 2023 Jun 9. PMID: 37294770; PMCID: PMC10256160.
