The normal lung cell line BEAS-2B and lung cancer cell line SPC-A-1 were purchased from The American Type Culture Collection (Manassas, VA, USA). Other lung cancer cell lines A549, H1975, H-125 and 95D were obtained from The Cell Bank of Chinese Academy of Sciences (Shanghai, China). All cells were cultured in Dulbecco's modified Eagle's medium (DMEM) purchased from Gibco (Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplied with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.) at 37°C. A FER1L4 expression plasmid was constructed using a pcDNA 3.1 vector by Jie Li Biology (http://www.genebioseq.com/ , Shanghai, China) with Xho I and HindIII restriction enzymes. A549, 95D and BEAS-2B cells were transfected with 2 µg/ml plasmid using Lipofectamine® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.) for 48 h, according to the manufacturer's protocol. SF1670 (Selleck Chemicals, Shanghai, China), a specific inhibitor of phosphatase and tensin homolog (PTEN), was purchased to activate the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway and diluted in dimethyl sulfoxide to a concentration of 50 mM. The working concentration of SF1670 was 50 µM and the incubation time was 6 h at 37°C.
Sf1670
Manufactured by Selleck Chemicals
Sourced in China, United States
SF1670 is a laboratory equipment designed for mixing and stirring liquids and solutions. It features a variable speed control and a digital display to monitor the rpm. The equipment is suitable for use in various scientific and research applications that require controlled mixing or agitation of samples.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Beas 2b, by American Type Culture Collection (1 mentions)
Beas 2b, by Thermo Fisher Scientific (1 mentions)
Indole 3 carbinol, by Merck Group (1 mentions)
Methyllycaconitine, by Merck Group (1 mentions)
Anti il10r, by BioXCell (1 mentions)
Pam3csk4, by InvivoGen (1 mentions)
Amd3100, by Selleck Chemicals (1 mentions)
Hyclone, by GE Healthcare (1 mentions)
Mk 2206, by Selleck Chemicals (1 mentions)
Igf 1, by Thermo Fisher Scientific (1 mentions)
6 protocols using sf1670
1
FER1L4 Overexpression in Lung Cells
2
Neuroprotective Effects of PTEN Inhibition
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Nicotine, methyllycaconitine (MLA), a selective α7 nAChR blocker (Gowayed et al., 2019 (link); Papke and Horenstein, 2021 (link)), and a potent PTEN protector (Lee et al., 2019 (link)), indole-3-carbinol (I3C), were purchased from Sigma-Aldrich. The inhibitor of PTEN, SF1670, was purchased from Selleckchem. SF1670 and I3C were dissolved in dimethylsulfoxide (DMSO) first and diluted in saline solution, while nicotine and MLA were dissolved in saline solution directly. Rats had already received intraperitoneal injections of nicotine at 1 mg/kg and MLA at 1 mg/kg once daily for 1 week before surgery and continued receiving intraperitoneal injections of 1 mg/kg nicotine or MLA in sterile saline once daily for 3 week (Teng et al., 2019 (link)), whereas matched control rats received an equal volume of intraperitoneal saline. SF1670 (0.3 mg/kg; Li et al., 2011 (link)) and I3C (2 mg/kg; Lee et al., 2019 (link)) were injected intraperitoneally in nicotine-treated rats simultaneously until the date of surgery to observe the nociceptive effect of PTEN deficiency and analgesic effects for the prevention of PTEN degradation, respectively.
3
Murine BMDM Isolation and Stimulation
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Murine BMDMs were obtained as previously described (Xu et al., 2012 (link)) and maintained in DMEM (HyClone) supplemented with 10% FBS (Gibco) and 10% supernatant of L929 cell as conditioned medium providing macrophage colony-stimulating factor (M-CSF, identified as complete medium). Briefly, bone marrow cells were extracted and cultured with complete medium for 5 days to derive BMDMs. Cell culture grade LPS and Pam3CSK4 were purchased from Invivogen and were used at a concentration of 10 ng/ml unless otherwise specified. SF1670 were from Selleck. anti-IL10R, anti-IgG, and anti-TNF were purchased from BioXCell. anti-IL10R or anti-IgG was added 30 min prior to LPS stimulation and were present throughout LPS exposure.
4
Inhibition of miR-10b-5p Impairs Colony Formation
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SGC-7901 or MGC-803cells plated into 6-well plates with a density of 1000 cells/well were transfected with miR-10b-5p inhibitor, treated with 1 μmol/L Oroxin B (Selleck, Houston, TX, USA), treated with 2 μmol/L SF1670 (Selleck), or cultured with the supernatant of MGC-803 cells' culture medium. After 10 days culture, cells were fixed and stained with crystal violet. The number of colonies were counted under a microscope from three independent replicates.
5
Vascular Endothelial Cell Inhibitor Study
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VECs (EA.hy926; The Cell Bank of Type Culture Collection of the Chinese Academy of Sciences) were cultured in DMEM (HyClone; GE Healthcare Life Sciences), supplemented with 5% FBS, 100 U/ml penicillin and 100 µg/ml streptomycin. VECs were incubated for 30 min with small molecule inhibitors, including AMD3100 (inhibitor of CXCR4), MK-2206 [inhibitor of AKT serine/threonine kinase 1 (AKT)] and SF1670 [inhibitor of phosphatase and tensin homolog (PTEN)], all purchased from Selleck Chemicals at a concentration of 10 µM. To suppress SDF1, VECs were treated with anti-SDF1 antibody (100 µg/ml; cat. no. ab155090; Abcam) for 24 h.
6
Modulating FAM46C Expression in Prostate Cancer
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FAM46C (NM_017709.3) interference sequences (position 222-240: 5’-CCAGGGATTGCATGTCCTT-3’; position 308-326: 5’-GGACGAGGCAACTTTCCAA-3’ position; 1296-1314: 5’-GCAACTTCAGCAACTACTA-3’) and a scramble control shRNA (shNC, 5’-GCGTCACTCAACATACATA-3’) were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China) and cloned into pLKO.1 vector to knockdown FAM46C expression. FAM46C overexpression plasmids were constructed by cloning full-length human FAM46C into the lentiviral expression vector pLVX-Puro (Addgen, Cambridge, MA, USA). Recombinant vectors (1000 ng) along with the psPAX2 (100 ng) and pMD2G (900 ng) packaging plasmids were co-transfected into 293T cells using Lipofectamine 2000. At 48 h after transfection, the supernatant was collected and transduced into 22RV1 or DU145 cells in the absence or presence of PTEN inhibitor SF1670 (250 nM; Selleck, Houston, TX, USA) or AKT signaling agonist IGF-1 (100 ng/ml; Peprotech, Rocky Hill, NJ, USA). Cells with pLKO.1-shNC or blank pLVX-Puro transduction were used as negative control.
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