NSCLC A549 and NCI-H460 cells were purchased from the Chinese Academy of Sciences Committee on Type Culture Collection Cell Bank (Shanghai, China) .Cell lines have been tested and authenticated by the centre. These cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM, GIBCO, USA) supplemented with 10% fetal bovine serum (Gibco, Grand Island, NY) and 1% penicillin/streptomycin (Beyotime Institute of Biotechnology, Shanghai, China). Cells were maintained at 37 °C in a humidified 5% CO2 atmosphere. EGCG were purchased from Sigma-Aldrich (St. Louis, MO, USA). Nocodazole, MG132 and Leupeptin were obtained from Beyotime Institute of Biotechnology (Shanghai, China).
Nocodazole
Manufactured by Beyotime
Sourced in China
Nocodazole is a chemical compound used as a laboratory tool. It functions as a microtubule-depolymerizing agent, inhibiting the formation of microtubules during the cell cycle.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Epigallocatechin gallate (egcg), by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Beyotime (1 mentions)
Mg132, by Beyotime (1 mentions)
Leupeptin, by Beyotime (1 mentions)
Genistein, by Beyotime (1 mentions)
Chlorpromazine, by Solarbio (1 mentions)
Dynasore, by GLPBIO (1 mentions)
Amiloride, by Solarbio (1 mentions)
Facsaria 2 flow cytometer, by BD (1 mentions)
Paclitaxel, by Selleck Chemicals (1 mentions)
Trichostatin a tsa, by Merck Group (1 mentions)
Taxol, by Selleck Chemicals (1 mentions)
Pf 573228, by Beyotime (1 mentions)
Mdl 28170, by MedChemExpress (1 mentions)
Eca109, by Shanghai Cell Bank (1 mentions)
Chlorpromazine hydrochloride, by Merck Group (1 mentions)
6 protocols using nocodazole
1
NSCLC Cell Culture and Reagents
2
Evaluating Cellular Uptake of mRNA-1096
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To detect cellular uptake, mRNA-1096 was fluorescently labeled with MFP488. DC 2.4 and RD cells were seeded at a cell density of 1/3 million in 12-well plates and treated with the MFP488-labeled DNCA/CLD-mRNA-1096 (MFP488-labeled mRNA-1096 = 1 μg) on 80–90% confluency. After incubated for different time periods at 37 °C in a 5% CO2 environment, cells were collected, fixed in 4% paraformaldehyde, and washed with PBS (pH = 7.4) prior to analysis on a BD FACS Aria II flow cytometer.
To determine the route of cellular internalization of DNCA/CLD-mRNA-1096, DC 2.4 and RD cells (at 80–90% confluency) were preincubated with chemical inhibitors, Genistein (50 μM; Beyotime) [37 (link),38 (link)], Chlorpromazine (15 μg/ml; Solarbio) [38 (link)], Dynasore (50 μM; Glpbio) [39 (link)], Nocodazole (1.25 μg/ml; Beyotime) [40 (link)], Amiloride (100 μM; Solarbio) [41 (link)], for 30 min before transfected with the MFP488-labeled DNCA/CLD-mRNA-1096 (MFP488-labeled mRNA-1096 = 1 μg). After incubation for 3 h, cells were harvested, fixed in 4% paraformaldehyde, and washed with PBS before the flow cytometric analysis.
To determine the route of cellular internalization of DNCA/CLD-mRNA-1096, DC 2.4 and RD cells (at 80–90% confluency) were preincubated with chemical inhibitors, Genistein (50 μM; Beyotime) [37 (link),38 (link)], Chlorpromazine (15 μg/ml; Solarbio) [38 (link)], Dynasore (50 μM; Glpbio) [39 (link)], Nocodazole (1.25 μg/ml; Beyotime) [40 (link)], Amiloride (100 μM; Solarbio) [41 (link)], for 30 min before transfected with the MFP488-labeled DNCA/CLD-mRNA-1096 (MFP488-labeled mRNA-1096 = 1 μg). After incubation for 3 h, cells were harvested, fixed in 4% paraformaldehyde, and washed with PBS before the flow cytometric analysis.
3
RABV Infection Inhibition Assay
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The chemicals used to treat the RABV-infected N2a cells included the microtubule-depolymerizing drug nocodazole (Noco) (S1765; Beyotime, China), the microtubule-polymerizing drug paclitaxel (Taxol) (S1150; Selleckchem, USA), the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) (T1952; Sigma) and the HDAC inhibitor sodium butyrate (NaBut) (S1539; Beyotime). The N2a cells were infected with RABV for 4 h and then treated with the various drugs. The solvent dimethyl sulfoxide (DMSO) was used as a control.
4
Dissection of Cellular Mechanics
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The inhibitors of FAK (PF573228; Cat. #SC1099, Beyotime, Shanghai, China), CaMKⅡ (KN93; Cat. #HY-15465, MedChemExpress, New Jersey, USA), calpeptin (MDL28170; Cat. #HY-18236, MedChemExpress), ATPase [(-)-blebbistatin; Cat. #SF9087, Beyotime], actin polymers (Cytochalasin B; Cat. #HY-16928, MedChemExpress), mitosis (Nocodazole; Cat. #S1765, Beyotime), and Rho-associated coiled kinase (ROCK; Y-27632; Cat. #SC0326, Beyotime) were used to determine the involved mechanical components
[
13 (link)]
. Calcium chloride anhydrous (CaCl
2; Cat. #C1016, Sigma, Shanghai, China), potassium chloride (KCl; Cat. #10016308, Sinapharm Chemical Reagent Co., Ltd., Shanghai, China), and magnesium chloride hexahydrate (Sinapharm Chemical Reagent Co., Ltd.) were used to treat HepG2 cells with different final concentrations.
[
13 (link)]
. Calcium chloride anhydrous (CaCl
2; Cat. #C1016, Sigma, Shanghai, China), potassium chloride (KCl; Cat. #10016308, Sinapharm Chemical Reagent Co., Ltd., Shanghai, China), and magnesium chloride hexahydrate (Sinapharm Chemical Reagent Co., Ltd.) were used to treat HepG2 cells with different final concentrations.
5
Esophageal Squamous Cell Carcinoma Protocol
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The human esophageal squamous cell carcinoma cell lines Eca109 and TE13 were obtained from Shanghai Cell Bank (Chinese Academy of Sciences, Shanghai, China). The cells were cultured in RPMI 1640 (Hyclone, Thermo Scientific, MA) supplemented with 10% fetal bovine serum (Hyclone) at 37°C under a humidified atmosphere of 5% CO2. YM155 monobromide was purchased from SelleckChem (Houston, TX). For the in vitro experiments, YM155 was dissolved in DMSO and diluted in medium to a final DMSO concentration of ≤0.1%. For the in vivo experiments, YM155 was dissolved and diluted in saline immediately before administration. Mitotic inhibitor nocodazole was purchased from Beyotime institute of biotechnology (Shanghai, China).
6
Baicalein-Loaded Liposomes for Enhanced Cellular Uptake
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Baicalein (98% purity) was purchased from Nanjing Zelang Biological Technology Co. Ltd. Soybean phospholipids were purchased from Shanghai Tywei Pharmaceutical Co. Ltd. Ethylis oleas was purchased from Jiangxi Alpha High‐Tech Pharmaceutical Co., Ltd. Tween 80 was cgqzlied by Shanghai Macklin Biochemical Co., Ltd. Transcutol HP was kindly donated by Gattefossé. Ascorbic acid and tetrahydrofuran were cgqzlied by Sinopharm Chemical Reagent Co., Ltd. Minimum Essential Medium (MEM), fetal bovine serum, non‐essential amino‐acid, L‐glutamine, penicillin, streptomycin, and 0.25% trypsin were purchased from Thermo Fisher Scientific. Coumarin 6 (Cou‐6), methyl‐β‐cyclodextrin, chlorpromazine hydrochloride, genistein, and amiloride hydrochloride were purchased from Sigma‐Aldrich. Brefeldin A, monensin, nocodazole, and 4,6‐diamino‐2‐phenyl indole (DAPI) were purchased from Beyotime Biotechnology Co., Ltd. Bafilomycin A1 was cgqzlied by Meilunbio. 1,1′‐Dioctadecyl‐3,3,3′,3′‐tetramethylindotricarbocyanine iodide (DiR) was obtained from AAT Bioquest Inc. Tetramethylrhodaminyl (TRITC) phalloidin was purchased from Yeasen Biotechnology Co., Ltd.
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