Human renal tubular epithelial cells (HK-2) were obtained from BeNa Culture Collection. Cells were grown in RPMI-1640 medium (Beijing Solarbio Science & Technology Co., Ltd.) containing 10% FBS at 37 °C with 5% CO2. In addition, cells were treated by different doses of remifentanil (0.625, 1.25, and 2.5 μM; Yichang Humanwell Pharmaceutical Co., Ltd.) for 24 h as needed [22 (link),23 (link)]. Tunicamycin (TM; 0.5, 1, 5, and 10 μg/ml; Abcam) [45–48 (link)] was used as an ERS inducer to pretreat cells at 37 °C for 2 h before treatment with remifentanil.
Tunicamycin tm
Manufactured by Abcam
Sourced in United Kingdom
Tunicamycin (TM) is a molecule that inhibits the N-linked glycosylation process. It acts by preventing the first step of this process, which is the synthesis of N-acetylglucosamine-1-phosphate transferase. This inhibition leads to the accumulation of unfolded proteins in the endoplasmic reticulum, triggering the unfolded protein response.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Solarbio (1 mentions)
4 phenylbutyrate 4 pba, by Merck Group (1 mentions)
3 methyladenine 3 ma, by Merck Group (1 mentions)
Hepg2, by Shanghai Cell Bank (1 mentions)
Control sirna, by GenePharma (1 mentions)
Hep3b, by Shanghai Cell Bank (1 mentions)
Cell counting kit 8 cck 8, by BestBio (1 mentions)
Sk hep 1, by Shanghai Cell Bank (1 mentions)
Plc prf 5, by Shanghai Cell Bank (1 mentions)
Antibody against β actin, by ZSGB-BIO (1 mentions)
Ab109234, by Abcam (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Ab214430, by Proteintech (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Hk 2 cell, by American Type Culture Collection (1 mentions)
4 protocols using tunicamycin tm
1
Remifentanil and Tunicamycin Effects on Renal Cells
2
Investigating Hepatocellular Carcinoma Cell Lines
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The human HCC cell lines (PLC/PRF/5, Hep3B, LM3, SK-HEP-1, HepG2 and Huh7) and normal hepatic cell line (LO2) were obtained from the Shanghai Cell Bank (Chinese Academy of Sciences, Shanghai, China). Sorafenib was purchased from Aladdin (Shanghai, China). Tunicamycin (TM) was purchased from Abcam (Cambridge, UK). Cell Counting Kit-8 (CCK-8) was purchased from BestBio (Nanjing, China). 4-phenylbutyrate (4-PBA) and 3-methyladenine (3-MA) were purchased from Sigma (USA). Antibody against GRP78 was purchased from Biogot Biotechnology (USA). Antibody against ZNF263 was purchased from Atlas antibodies (Bromma, Sweden). Antibody against Beclin 1 and LC 3 were purchased from Abcam (Cambridge, UK). Antibody against XBP-1 s was purchased from CST (USA). Antibody against GAPDH was purchased from Bioss antibodies (Beijing, China). Antibody against β-actin was purchased from ZSGB-BIO Inc. (Beijing, China). The ZNF263 small interfering RNA (siRNA) and control siRNA were purchased from GenePharma (Shanghai, China).
3
Optimizing HK-2 Cell Culture for Kidney Research
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The HK-2 cells were obtained from the American Type Culture Collection (ATCC, USA). DMEM/F12 was obtained from HyClone (SH30023.01, USA). Fetal bovine serum was obtained from Gibco (FBS-CBT, USA). Tunicamycin (TM) was acquired from Abcam (ab120296, UK). Paricalcitol was obtained from USP (1499414, USA). ATF4 plasmid and siRNA were constructed in Guangzhou Ribo Biotechnology Co, Ltd. lipofectamine3000 was acquired from Invitrogen (L3000-018, Guangzhou, China). The following antibodies were used in the experiments: anti-VDR (Abcam, ab109234), anti-ATF4 (Abcam, ab184909), anti-BiP (Proteintech,11587-1-AP, Wuhan, China), anti-CHOP (Proteintech,15204-1-AP), anti-cleaved-caspase3 (Abcam, ab214430) and anti-β-actin (Proteintech,20536-1-AP).
4
Arabidopsis Stress Response Assay
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The seeds of Arabidopsis were germinated and grew on half-strength MS medium with or without 0.3 µg/mL tunicamycin (TM, Abcam) for 7 days. Two-week seedlings were treated with 5 µg/mL TM for 6 h or with 0.25% DMSO (control) for RT-PCR analysis. The seeds were germinated and grew on half-strength MS medium with 120 mM NaCl for 12 days to cause stress response.
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