Human SCLC cells (NCI-H1688) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were grown in RPMI 1640 medium (Gibco, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (Gibco, USA). DNA sequences of shRNAs targeting TCONS_00020615 were subcloned into the pLKO.1-TRC lentivirus vector. The sequences of shRNAs against TCONS_00020615 were 5'-CAGCCCACAAATAAACTGGTA-3'; 5′-GTGCAGGTTGCATTTACTTAT-3'; 5′-AACCAACAGCTTTCAAAGTAA-3'; shRNA directed against GFP (target sequence: 5'-GCAAGCTGACCCTGAAGTTCAT-3') was used as the control scrambled shRNA (shCtrl). Cells were infected with lentivirus carrying specific shRNA in the presence of polybrene (8 μg/mL, sigma) for 24 h. The medium was replaced with fresh medium and the cells were cultured for 48 h. Then the cells were screened with selection medium containing 2.5 μg/mL puromycin until there was no live cell left in the mock group.
Nci h1688
Manufactured by American Type Culture Collection
Sourced in United States
The NCI-H1688 is a cell line derived from human lung cancer tissue. It is a type of adherent cell line that can be used for various cell culture and research applications. The core function of this cell line is to provide a model system for the study of lung cancer biology and potential therapeutic interventions.
Automatically generated - may contain errors
Lab products found in correlation
Nci h446, by American Type Culture Collection (3 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Polybrene, by Merck Group (1 mentions)
Puromycin, by Merck Group (1 mentions)
Nci h69, by American Type Culture Collection (1 mentions)
Nci h69ar, by American Type Culture Collection (1 mentions)
Nci h146, by American Type Culture Collection (1 mentions)
Humidified incubator, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Nci h520, by American Type Culture Collection (1 mentions)
Nci h460, by American Type Culture Collection (1 mentions)
Nci h441, by American Type Culture Collection (1 mentions)
Nci h661, by American Type Culture Collection (1 mentions)
Nci h1993, by American Type Culture Collection (1 mentions)
Mrc 5, by American Type Culture Collection (1 mentions)
Nci h226, by American Type Culture Collection (1 mentions)
Cor l23, by American Type Culture Collection (1 mentions)
8 protocols using nci h1688
1
Knockdown of lncRNA TCONS_00020615 in SCLC
2
Culturing Small Cell Lung Cancer Cell Lines
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The human SCLC cell line NCI-H69 and the drug-resistant sublines NCI-H69AR, NCI-H446, NCI-H146, and NCI-H1688 were purchased from the American Type Culture Collection (ATCC; Rockville, MD, USA) and maintained in RPMI 1640 medium (HyClone, Logan, UT, USA) with 10 % fetal calf serum (HyClone) in an incubator at 37 °C with 5 % CO2. The H69AR subline was maintained in a 5 μg/L final concentration of doxorubicin (Jiangshu, China) and transferred to drug-free media for at least 2 weeks before any experiment.
3
Cell Culture of NCI-H446 and NCI-H1688
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The human SCLC cell lines NCI-H446 and NCI-H1688 were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA). The cells were cultured in RPMI 1640 (Invitrogen, Beijing, China) supplemented with 10% fetal bovine serum (FBS, Invitrogen, Carlsbad, CA, USA). All cells were incubated at 37°C in a humidified incubator (Thermo Electron Corp, New Castle, DE) containing 5% CO2.
4
Diverse Human Lung Cells Authenticity
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NL20 human bronchial epithelial cells and NCI-H460, NCI-H661, NCI-H1993, NCI-H441, NCI-H520, NCI-H1688, and A549 human lung cancer cell lines were purchased from the American Type Culture Collection (ATCC) and authenticated by ATCC based on their DNA profiles, cytogenetic analyses, and isoenzymology. These cells were cultured according to ATCC's protocols and passaged for fewer than 6 months after resuscitation. CL1-5 cells were established by Chu et al. 64 (link) and routinely verified (based on their growth, morphology, and lack of mycoplasma contamination) in our laboratory. Human normal nasal mucosal epithelial (NNM) cells were a primary culture derived from a nasal polyp 65 (link), and were grown in DMEM.
5
Establishing Lung Cancer Cell Line Cultures
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Five cell lines were purchased from American Type Culture Collection (ATCC; Rockville, MD, USA), namely three subtypes of human non-small cell lung cancer (NSCLC) consisting of A549 (adenocarcinoma), COR-L23 (large cell carcinoma), and NCI-H226 (squamous cell carcinoma); human small cell lung cancer (SCLC) in the form of NCI-H1688; and human normal lung fibroblasts as MRC-5. All cancer cell lines were maintained in RPMI-1640 supplemented with 10% FBS, and MRC-5 cells were cultured in MEM supplemented with 10% FBS, 1 mM nonessential amino acid, and 1% sodium pyruvate at 37°C in 5% CO2 incubator.
6
Characterization of TP53 Mutant Cancer Cell Lines
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HDQ-P1 cells were purchased from the German Collection of Microorganisms and Cell Cultures, and HCT116, SW900, NCI-H1688, HCC1937 and NCI-H1299 cells from American Type Culture Collection. NCI-H1299 cells stably expressing p53 R213X (TGA) were generated in our laboratory. HCT116 cells are homozygous for wild type TP53 and the following cancer cell lines harbor homozygous nonsense mutations in the TP53 gene: HDQ-P1 (R213X), SW900 (Q167X), NCI-H1688 (Q192X), HCC1937 (R306X) [14 (link)]. HDQ-P1 and HCT116 cells were cultured in high glucose Dulbecco’s modified Eagle medium (DMEM, Sigma-Aldrich) supplemented with 10% (vol/vol) fetal bovine serum (FBS, Sigma-Aldrich) and 1× antibiotic–antimycotic (Gibco/Thermo Fisher Scientific) at 37°C and 5% (vol/vol) CO2. SW900, NCI-H1688, HCC1937, and NCI-H1299 cells were cultured in RPMI-1640 medium (Sigma-Aldrich) supplemented with 10% (vol/vol) FBS (Sigma-Aldrich) and 1× antibiotic–antimycotic (Gibco/Thermo Fisher Scientific) at 37°C and 5% (vol/vol) CO2.
7
Culturing Cancer Cell Lines and Isolating Primary Myeloma Cells
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Cervical cancer cell line HeLa, human embryonic kidney cell line HEK293T, multiple myeloma cell lines MM1.S and RPMI8226, and small cell lung cancer cell line NCI-H1688 were obtained from American Type Culture Collection (ATCC). Multiple myeloma CAG cells (Borset et al., 2000 (link)) were a kind gift from Dr. Joshua Epstein (University of Arkansas for Medical Sciences, Little Rock, AK, United States). MM1.S, RPMI8226, CAG, and NCI-H1688 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium. HeLa and HEK293T cells were cultured in Dulbecco′s Modified Eagle’s Medium (DMEM; HyClone). Growth medium was supplemented with 10% fetal bovine serum (FBS; Gibco and Lonsera), 100 U/ml penicillin, and 100 μg/ml streptomycin (Gibco).
Bone marrow specimens were obtained from deidentified multiple myeloma patients at the Weill Cornell Medicine under informed consent as part of an Institutional Review Board approved study. CD138+ primary myeloma cells were isolated from bone marrow and co-cultured with a layer of HS-5 cells and cytokines as previously described (Huang et al., 2012 (link)).
Bone marrow specimens were obtained from deidentified multiple myeloma patients at the Weill Cornell Medicine under informed consent as part of an Institutional Review Board approved study. CD138+ primary myeloma cells were isolated from bone marrow and co-cultured with a layer of HS-5 cells and cytokines as previously described (Huang et al., 2012 (link)).
8
In Vitro and In Vivo Evaluation of Novel Anticancer Agents
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NCI-H1688, SK-OV-3, MDA-MB-231,
HT-29, HepG2, and NCI-H446 cell lines were purchased from the American
Type Culture Collection (Rockville, MD, USA). McCoy’s 5A, DMEM,
RPMI-1640 medium, and fetal bovine serum (FBS) were purchased from
Gibco (Gaithersburg, MD, USA); the absorbance and fluorescence were
measured using a Victor Nivo Microplate Reader (Perkin Elmer, German);
a Cell Cycle Assay Kit was purchased from US Everbright Inc. (Suzhou,
China); a tubulin polymerization assay kit (BK011P) was purchased
from Cytoskeleton Inc. (Denver, CO, USA); Western blot analyses were
performed with primary antibodies against the following antigens:
mouse anti-phospho-c-Src (1:100; Santa Cruz Cat# sc-81521), mouse
anti-c-Src (1:200, Santa Cruz Cat# sc-8056), anti-paxillin (pY118)
antibody (1:500; Cohesion Biosciences Cat# CPA3307), anti-paxillin
(1:500; Santa Cruz Cat# sc-365379), rabbit anti-GAPDH (1:1000; Cell
Signaling Technology Cat# 2118), anti-rabbit IgG, HRP-linked antibody
(1:4000; Cell Signaling Technology Cat# 7074), anti-mouse IgGκ,
and HRP-linked antibody (1:4000; Santa Cruz Cat# sc-516102). Five-week-old
female non-obese diabetic (NOD)/severe-combined immunodeficient mouse
(SCID) and male Sprague–Dawley rats were obtained from Weitonglihua
(Beijing, China).
HT-29, HepG2, and NCI-H446 cell lines were purchased from the American
Type Culture Collection (Rockville, MD, USA). McCoy’s 5A, DMEM,
RPMI-1640 medium, and fetal bovine serum (FBS) were purchased from
Gibco (Gaithersburg, MD, USA); the absorbance and fluorescence were
measured using a Victor Nivo Microplate Reader (Perkin Elmer, German);
a Cell Cycle Assay Kit was purchased from US Everbright Inc. (Suzhou,
China); a tubulin polymerization assay kit (BK011P) was purchased
from Cytoskeleton Inc. (Denver, CO, USA); Western blot analyses were
performed with primary antibodies against the following antigens:
mouse anti-phospho-c-Src (1:100; Santa Cruz Cat# sc-81521), mouse
anti-c-Src (1:200, Santa Cruz Cat# sc-8056), anti-paxillin (pY118)
antibody (1:500; Cohesion Biosciences Cat# CPA3307), anti-paxillin
(1:500; Santa Cruz Cat# sc-365379), rabbit anti-GAPDH (1:1000; Cell
Signaling Technology Cat# 2118), anti-rabbit IgG, HRP-linked antibody
(1:4000; Cell Signaling Technology Cat# 7074), anti-mouse IgGκ,
and HRP-linked antibody (1:4000; Santa Cruz Cat# sc-516102). Five-week-old
female non-obese diabetic (NOD)/severe-combined immunodeficient mouse
(SCID) and male Sprague–Dawley rats were obtained from Weitonglihua
(Beijing, China).
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